TEMPLATE ABHÄNGIGE LIGIERUNG MIT PNA-DNA CHIMERISCHEN SONDEN

PEPTIDNUCLEINSÄUREN

BY MEANS OF NUKLEINSÄUREANALOGS INDUCED TRANSKRIPTION OF DOPPELSTRÄNGIGER DNA

Compositions of solvents and high concentrations of nucleic acid analogs

PEPTIDE NUCLEIC ACIDS

A novel class of compounds, known as peptide nucleic acids, bind complementary ssDNA and RNA strands more strong- ly than a corresponding DNA. The peptide nucleic acids generally comprise ligands such as naturally occurring DNA bases att- ached to a peptide backbone through a suitable linker.

PEPTIDE NUCLEIC ACIDS

A novel class of compounds, known as peptide nucleic acids, bind complementary ssDNA and RNA strands more strong-ly than a corresponding DNA. The peptide nucleic acids generally comprise ligands such as naturally occurring DNA bases att-ached to a peptide backbone through a suitable linker.

USES OF NUCLEIC ACID ANALOGUES IN THE INHIBITION OF NUCLEIC ACID AMPLIFICATION

Nucleic acid analogues of the kind described in PCT/EP92/01220 (PNA's) which hybridise strongly to nucleic acids are used to inhibit nucleic acid amplification procedures such as PCR. False positives in subsequent PCR assays are prevented by hybridising a PNA to PCR amplification products. Assays capable of discriminating between single base pair mutants are con- ducted by using a PNA hybridising to one of the two allelic forms to inhibit a PCR amplification of that form selectively. Asym- metric PCR amplifications are carried out by starting a PCR symmetricaly usi ng like quantities of forward and reverse primers and once the amplification is established disabling one primer by hybridisin g a PNA thereto.

BINARY PROBE, CLIP COMPOSITION AND PROCEDURE FOR THE GOAL HYBRIDIZING PROOF

Peptide nucleic acids having enhanced binding affinity, sequence specificity and solubility

Use of nucleic acid analogues in the inhibition of nucleic acid amplification

NOVEL PEPTIDE NUCLEIC ACIDS

A novel class of compounds, known as peptide nucleic acids, bind complementary ssDNA and RNA strands more strongly than a corresponding DNA. The peptide nucleic acids generally comprise ligands such as naturally occurring DNA bases attached to a peptide backbone through a suitable linker.

Use of nucleic acid analogues in diagnostics and analytical procedures

Methods of capture, recognition, detection, identification or quantitation of nucleic acids and diagnostics uses generally are described in which are used: (a) a peptide nucleic acid (PNA) comprising a polyamide backbone bearing a plurality of ligands at respective spaced locations along said backbone, said ligands being each independently naturally occurring nucleobases, non-naturally occurring nucleobases or nucleobase-binding groups, each said ligand being bound directly or indirectly to a nitrogen atom in said backbone, and said ligand bearing nitrogen atoms mainly being separated from one another in said backbone by from 4 to 8 intervening atoms; or (b) a nucleic acid analogue capable of hybridizing to a nucleic acid of complementary sequence to form a hybrid which is more stable against denaturation by heat than a hybrid between the conventional deoxyribonucleotide corresponding to said analogue and said nucleic acid; or (c) a nucleic acid analogue capable of hybridizing to a double ...

SYNTHONE FÜR DIE SYNTHESE UND DAS ENTSCHÜTZEN VON PEPTIDISCHEN NUCLEINSÄUREN UNTER MILDEN BEDINGUNGEN.

LINKED PEPTID NUCLEIC ACIDS

STRUCTURE OF HIGHER ORDER AND CONNECTION OF PEPTIDNUKLEINSÄUREN

PEPTID NUCLEIC ACID KONJUGATE ONE

USE OF NUCLEIC ACID SIMILAR ONE IN INHIBITION OF NUCLEIC ACID AMPLIFICATION

COMPOSITIONS OF SOLVENTS AND HIGHLY CONCENTRATED ONE NUCLEIC ACID ANALOGUES

EXTENSION OF A PNA-DNA CHIMÄRE AT YOUR 3 ' END OF MEANS OF A POLYMERASE

TEMPLATE DEPENDENT ONES LIGIERUNG WITH PNA-DNA CHIMERI PROBES

Binary probe and clamp composition and methods for target hybridization detection

Binary probe and clamp compositions conduct methods for target hybridization detection. Where the probe is a substrate for exonuclease cleavage, the composition provides quantitation and detection of PCR products, by real-time and end-point measurements. Where the probe is an amplification primer, the composition provides an improved method for labelling and detection of PCR products. Probes and clamps may be labelled with fluorescent dyes, quenchers, hybridization-stabilizing moieties, chemiluminescent dyes, and affinity ligands. Clamps may be nucleic acid analogs, such as 2-aminoethylglycine PNA.

Nucleic acid analogue-induced transcription of RNA from a double-stranded DNA template

RNA is transcribed from a double-stranded DNA template by forming a complex by hybridising to the template at a desired transcription initiation site one or more oligonucleic acid analogues, preferably of the PNA (peptide/nucleic acid) type, capable of forming a transcription initiation site with the DNA and exposing the complex to the action of a DNA dependant RNA polymerase in the presence of nucleoside triphosphates. Equal length transcripts may be obtained by placing a block to transcription downstream from the initiation site or by cutting the template at such a selected location. The initiation site may be formed by displacement of one strand of the DNA locally by the PNA hybridisation.

ASYNCHRONOUS PCR PRIMING

DOPPELSTRÄNGIGE PEPTIDENUKLEINSÄUREN

PEPTIDNUKLEINSÄUREN

PEPTIDNUKLEINSÄUREN WITH INCREASED CONNECTION AFFINITY, SEQUENCE SPECIFICITY AND SOLUBILITY

Compositions of solvents and high concentrations of nucleic acid analogs

Binary probe and clamp composition

Binary probe and clamp compositions conduct methods for target hybridization detection. Where the probe is a substrate for exonuclease cleavage, the composition provides quantitation and detection of PCR products, by real-time and end-point measurements. Where the probe is an amplification primer, the composition provides an improved method for labelling and detection of PCR products. Probes and clamps may be labelled with fluorescent dyes, quenchers, hybridization-stabilizing moieties, chemiluminescent dyes, and affinity ligands. Clamps may be nucleic acid analogs, such as 2-aminoethylglycine PNA.

Peptide nucleic acids

A novel class of compounds known as peptide nucleic acids, bind complementary DNA and RNA strands, and generally do so more strongly than the corresponding DNA or RNA strands while exhibiting increased sequence specificity and solubility. The peptide nucleic acids comprise ligands selected from a group consisting of naturally-occurring nucleobases and non-naturally-occurring nucleobases, including 2,6-diaminopurine, attached to a polyamide backbone, and contain alkyl amine side chains.

NUCLEIC ACID AMPLIFICATION

Novel peptide nucleic acids

NUCLEIC ACID ANALOGUE INDUCED TRANSCRIPTION OF DOUBLE STRANDED DNA

RNA is transcribed from a double stranded DNA template by forming a complex by hybridising to the template at a desired transcription initiation site one or more oligonucleic acid analogues of the PNA type capable of forming a transcription initiation site with the DNA and exposing the complex to the action of a DNA dependant RNA polymerase in the presence of nucleoside triphosphates. Equal length transcripts may be obtained by placing a block to transcription downstream from the initiation site or by cutting the template at such a selected location The initiation site is formed by displacement of one strand of the DNA locally by the PNA hybridisation.

Verwendung von Nuklein-säure Analoge in der Inhibition von Nuklein-säure Amplifikation

Asynchronous primed pcr

SYSTEM AND METHOD FOR IMPROVED PROCESSING OF NUCLEIC ACIDS FOR PRODUCTION OF SEQUENCABLE LIBRARIES

An embodiment of an adaptor element for efficient target processing is described that comprises a semi-complementary double stranded nucleic acid adaptor comprising a non- complementary region and a complementary region, where the non-complementary region comprises a first amplification primer site and a second amplification primer site and the complementary region comprises a sequencing primer site and one or more inosine species.

Nucleic Acid Analogue Induced Transcription of Double Stranded DNA

ZUSAMMENSETZUNGEN VON LÖSUNGSMITTELN UND HOCHKONZENTRIERTE NUKLEINSÄUREANALOGA

A new achiral linker reagent for the incorporation of multiple amino groups into oligonucleotides

DOUBLE-STRANDED PEPTIDE NUCLEIC ACIDS

A novel class of compounds, known as peptide nucleic acids, form double-stranded structures with one another and with ssDNA. The peptide nucleic acids generally comprise ligands such as naturally occurring DNA bases attached to a peptide backbone through a suitable linker.

Linked peptide nucleic acids

Peptide nucleic acids having enhanced binding affinity, sequence specificity and solubility

Peptide nucleic acid conjugates

Template-dependent ligation with PNA-DNA chimeric probes

Methods for isolating one strand of a double-stranded nucleic acid

Polymerase extension at 3' terminus of pna-dna chimera

Peptide nucleic acids having 2,6-diaminopurine nucleobases

A novel class of compounds, known as peptide nucleic acids, bind complementary DNA and RNA strands more strongly than a corresponding DNA strand, and exhibit increased sequence specificity and binding affinity. The peptide nucleic acids of the invention comprise ligands selected from a group consisting of naturally-occurring nucleobases and non-naturally-occurring nucleobases attached to a polyamide backbone. Some PNAs of the invention also contain C1-C8 alkylamine side chains.

HIGHER ORDER STRUCTURE AND BINDING OF PEPTIDE NUCLEIC ACIDS

Peptide nucleic acids and analogues of peptide nucleic acids are used to form duplex, triplex, and other structures with nucleic acids and to modify nucleic acids. The peptide nucleic acids and analogues thereof also are used to modulate protein activity through, for example, transcription arrest, transcription initiation, and site specific cleavage of nucleic acids.

THE USE OF NUCLEIC ACID ANALOGUES IN DIAGNOSTICS AND ANALYTICAL PROCEDURES

THE USE OF NUCLEIC ACID ANALOGUES IN DIAGNOSTICS AND ANALYTICAL PROCEDURES Methods of capture, recognition, detection, identification or quantitation of nucleic acids and diagnostics uses generally are described in which are used (a) a peptide nucleic acid (PNA) comprising a polyamide backbone bearing a plurality of ligands at respective spaced locations along said backbone, said ligands being each independently naturally occurring nucleobases, non-naturally occurring nucleobases or nucleobase-binding groups, each said ligand being bound directly or indirectly to a nitrogen atom in said backbone, and said ligand bearing nitrogen atoms mainly being separated from one another in said backbone by from 4 to 8 intervening atoms; (b) a nucleic acid analogue capable of hybridising to a nucleic acid of complementary sequence to form a hybrid which is more stable against denaturation by heat than a hybrid between the conventional deoxyribonucleotide corresponding to said analogue and said nucleic ...

PEPTIDE NUCLEIC ACIDS HAVING ENHANCED BINDING AFFINITY, SEQUENCE SPECIFICITY AND SOLUBILITY

A novel class of compounds, known as peptide nucleic acids, bind complementary DNA and RNA strands more strongly than a corresponding DNA strand, and exhibit increased sequence specificity and solubility. The peptide nucleic acids comprise ligands selected from a group consisting of naturally-occurring nucleobases and non-naturally-occurring nucleobases attached to a polyamide backbone, and contain C1-C8 alkylamine side chains. Methods of enhancing the solubility, binding affinity and sequence specificity of PNAs are provided.

Nucleic acid analogue-induced transcription of RNAfrom a double-stranded DNA template

Methods for isolating one strand of a double-stranded nucleic acid

USES OF NUCLEIC ACID ANALOGUES IN THE INHIBITION OF NUCLEIC ACID AMPLIFICATION

Nucleic acid analogues of the kind described in PCT/EP92/01220 (PNA's) which hybridise strongly to nucleic acids are used to inhibit nucleic acid amplification procedures such as PCR. False positives in subsequent PCR assays are prevented by hybridising a PNA to PCR amplification products. Assays capable of discriminating between single base pair mutants are con-ducted by using a PNA hybridising to one of the two allelic forms to inhibit a PCR amplification of that form selectively. Asym-metric PCR amplifications are carried out by starting a PCR symmetricaly using like quantities of forward and reverse primers and once the amplification is established disabling one primer by hybridising a PNA thereto.

MITTELS NUKLEINSÄUREANALOGS INDUZIERTE TRANSKRIPTION VON DOPPELSTRÄNGIGER DNA

VERLÄNGERUNG EINER PNA-DNA CHIMÄRE AN IHREM 3' ENDE MITTELS EINER POLYMERASE

Multipartite high-affinity nucleic acid probes

The use of nucleic acid analogues in diagnostics and analytical procedures

VERKNÜPFTE PEPTID-NUKLEINSÄUREN

Peptidnukleinsäuren

Novel peptide nucleic acids

Binary probe and clamp composition and methods for target hybridization detection

VERWENDUNG VON NUKLEINSÄUREANALOGA IN DIAGNOSTISCHEN UND ANALYTISCHEN VERFAHREN

DETECTION METHOD USING DISSOCIATED ROLLING CIRCLE AMPLIFICATION

Use of nucleic acid analogues in the inhibition of nucleic acid amplification

Template-dependent ligation with PNA-DNA chimeric probes

DOPPELSTRÄNGIGE PEPTIDENUKLEINSÄUREN

VERLÄNGERUNG EINER PNA-DNA CHIMÄRE AN IHREM 3' ENDE MITTELS EINER POLYMERASE

ASYNCHRONES PCR PRIMING

STRUKTUR HÖHERER ORDNUNG UND BINDUNG VON PEPTIDNUKLEINSÄUREN

Binary probe and clamp composition and methods for target hybridization detection

BINÄRSONDE, KLAMMERZUSAMMENSETZUNG UND VERFAHREN ZUM ZIELHYBRIDISIERUNGSNACHWEIS

Paired end sequencing

The present invention provides for a method of preparing a target nucleic acid fragments to produce a smaller nucleic acid which comprises the two ends of the target nucleic acid. Specifically, the invention provides cloning and DNA manipulation strategies to isolate the two ends of a large target nucleic acid into a single small DNA construct for rapid cloning, sequencing, or amplification.

PEPTID-NUKLEINSÄURE-KONJUGATE

BINARY PROBE AND CLAMP COMPOSITION AND METHOD, FOR TARGET HYBRIDIZATION DETECTION

PROBLEM TO BE SOLVED: To provide binary probe and clump compositions useful for performing methods for target hybridization detection. SOLUTION: Where the probe is a substrate for exonuclease cleavage, the composition provides quantitation and detection of PCR products, by real-time and end-point measurements. Where the probe is an amplification primer, the composition provides an improved method for labeling and detection of PCR products. Probes and clamps may be labeled with fluorescent dyes, quenchers, hybridization-stabilizing moieties, chemiluminescent dyes, and affinity ligands. Clamps may be nucleic acid analogs, such as 2-aminoethylglycine PNA. COPYRIGHT: (C)2009,JPO&INPIT ...

MITTELS NUKLEINSÄUREANALOGS INDUZIERTE TRANSKRIPTION VON DOPPELSTRÄNGIGER DNA

VERFAHREN UND TESTSYSTEM ZUR HYBRIDISIERUNGSANALYSE UNTER VERWENDUNG VON PEPTIDNUKLEINSÄURE-SONDEN

Verwendung von Nuklein-säure Analoge in der Inhibition von Nuklein-säure Amplifikation

PEPTIDNUKLEINSÄUREN MIT ERHÖHTER BINDUNGSAFFINITÄT, SEQUENZSPEZIFITÄT UND LÖSLICHKEIT

VERWENDUNG VON NUKLEINSÄUREANALOGA IN DIAGNOSTISCHEN UND ANALYTISCHEN VERFAHREN

Modulation of cellular transcription factor activity

A novel class of compounds, known as peptide nucleic acids, form double-stranded structures with one another and with ssDNA. The peptide nucleic acids generally comprise ligands such as naturally occurring DNA bases attached to a peptide backbone through a suitable linker.

BINÄRSONDE, KLAMMERZUSAMMENSETZUNG UND VERFAHREN ZUM ZIELHYBRIDISIERUNGSNACHWEIS

PEPTIDNUKLEINSÄUREN MIT ERHÖHTER BINDUNGSAFFINITÄT, SEQUENZSPEZIFITÄT UND LÖSLICHKEIT

Multipartite high-affinity nucleic acid probes

The invention provides a collection of probes useful for hybridizing to a target nucleic acid. The probes associate with each other, binding with high affinity to the target nucleic acid, to form three-way junctions and other complexes. At least one of the probes in each collection includes a nucleic acid analog. Methods using the probes in hybridization and as primers are also provided.

VERFAHREN UND TESTSYSTEM ZUR HYBRIDISIERUNGSANALYSE UNTER VERWENDUNG VON PEPTIDNUKLEINSÄURE-SONDEN

ZUSAMMENSETZUNGEN VON LÖSUNGSMITTELN UND HOCHKONZENTRIERTE NUKLEINSÄUREANALOGA

Template Abhängige Ligierung mit PNA-DNA chimerischen Sonden

Peptidnukleinsäuren

Linked peptide nucleic acids

Novel peptide nucleic acids and novel linked peptide nucleic acids, form triple stranded structures with nucleic acids. The peptide nucleic acids include ligands such as naturally occurring nucleobases attached to the peptide backbone through a suitable linker. Other nucleobases including C-pyrimidines and iso-pyrimidines can be used as the ligands in Hoogsteen strands to increase binding affinity. Two peptide nucleic acid strands are joined together with a linker to form a bis-peptide nucleic acid. The individual strands of the peptide nucleic acids in the bis compounds can be oriented either parallel or antiparallel to each other.

STRUKTUR HÖHERER ORDNUNG UND BINDUNG VON PEPTIDNUKLEINSÄUREN

Peptide nucleic acid synthons

A novel class of compounds, known as peptide nucleic acids, bind complementary ssDNA and RNA strands more strongly than a corresponding DNA. The peptide nucleic acids generally comprise ligands such as naturally occurring DNA bases attached to a peptide backbone through a suitable linker.

DOPPELSTRÄNGIGE PEPTIDENUKLEINSÄUREN

PEPTID-NUKLEINSÄURE-KONJUGATE

Nucleic acid amplification

Disclosed are compositions and a method for amplification of nucleic acid sequences of interest. The disclosed method generally involves replication of a complex nucleic acid sample such as genomic samples using one, a few, or more primers such that, during replication, the replicated strands are displaced from the nucleic acid molecules in the sample by strand displacement replication of another replicated strand. It was discovered that highly complex nucleic acid samples can be efficiently amplified using only one or a few primers having specific nucleic acid sequences. The one or few primers are complementary to nucleic acid sequences distributed throughout nucleic acid in the sample.

NUCLEIC ACID ANALOGUE, DIAGNOSTIC AGENT, AND USE OF THEM IN ANALYZING METHOD

PROBLEM TO BE SOLVED: To provide some nucleic acid analogues in order to perform capture, recognition, detection, identification or quantification about one or more chemically or microbiologically individual units for example. SOLUTION: The nucleic acid analogue with a new structure having characteristics that have not been conventionally known such as a characteristic to form a hybrid with a conventionally known nucleic acid having a complementary arrangement and to make stability in the Tm value of the hybrid higher than the hybrid formed by the conventionally known nucleic acid having an equivalent arrangement and/or a characteristic to make selectivity to the complementary arrangement compared with an arrangement relating to a degree of mismatching higher than the known conventional nucleic acid having the equivalent arrangement, and the method of using the nucleic acid analogues for diagnostic agents or analysis are provided. COPYRIGHT: (C)2009,JPO&INPIT ...

Detection method using dissociated rolling circle amplification

Disclosed are compositions and methods for detecting small quantities of analytes such as proteins and peptides. The method involves associating a DNA circle with the analyte and subsequent release and rolling circle replication of the circular DNA molecule. In the method, an amplification target circle is associated with analytes using a conjugate of the circle and a specific binding molecule that is specific for the analyte to be detected. Amplification target circles not associated with the proteins are removed, the amplification target circles that are associated with the proteins are decoupled from the specific binding molecule and amplified by rolling circle amplification. The amplification is isothermic and can result in the production of a large amount of nucleic acid from each primer. The amplified DNA serves as a readily detectable signal for the analytes.

NEW PEPTIDE NUCLEIC ACID

PROBLEM TO BE SOLVED: To obtain a new peptide nucleic acid comprising a specific peptide nucleic acid, having an action killing cells or virucidal action by modulating the expression of a gene, useful as a medicine composition for treating, diagnosing and preventing diseases, etc., as a gene targeting agent. SOLUTION: This new peptide nucleic acid is shown by formula I [(n) is at least 2; L1 to Ln are each H, OH, a 1-4C alkanoyl, a nucleus base, an aromatic part, a DNA intercalator, a nucleus base binding group, a heterocyclic ring part, a reporter ligand or the like; C1 to Cn are each a group of the formula (CR6R7)y (R6 is H; R7 is a side chain of a naturally occurring α amino acid; or R6 and R7 are each H, a 2-6C alkyl, an aryl, a heteroaryl, hydroxy, a 1-6C alkoxy or the like; (u) is 0 or 1-10) or the like; D1 to Dn are each a group of the formula (CR6R7)z ((z) is 0 or 1-10); G1 to Gn-1 are each a group of the formula NR3CO (R3 is H, a 1-9C alkyl, OH, an alkoxy or the like) or the like ...

PNA-DNA chimeric probe arrays and methods of use

The invention provides methods, kits, and compositions for ligation of PNA-DNA chimeric probes and oligonucleotides when they are hybridized adjacently to template nucleic acids using ligases and ligation reagents. Structural requirements of the chimeras for ligation include 5 to 15 contiguous PNA monomer units, 2 or more contiguous nucleotides, and a 3' hydroxyl or 5' hydroxyl terminus. The chimera and/or oligonucleotide may be labelled with fluorescent dyes or other labels. The methods include, for example, oligonucleotide-ligation assays (OLA) and single nucleotide polymorphism detection.

Binary probe and clamp composition and methods for a target hybridization detection

Binary probe and clamp compositions conduct methods for target hybridization detection. Where the probe is a substrate for exonuclease cleavage, the composition provides quantitation and detection of PCR products, by real-time and end-point measurements. Where the probe is an amplification primer, the composition provides an improved method for labelling and detection of PCR products. Probes and clamps may be labelled with fluorescent dyes, quenchers, hybridization-stabilizing moieties, chemiluminescent dyes, and affinity ligands. Clamps may be nucleic acid analogs, such as 2-aminoethylglycine PNA.

NUCLEIC ACID ANALOG, DIAGNOSTIC AGENT AND METHOD FOR USING THE SAME IN ANALYSIS METHOD

PROBLEM TO BE SOLVED: To provide a nucleic acid analog for performing capture, recognition, detection, identification or quantification about chemically or microbiologically individual units. SOLUTION: The nucleic acid analogues provided are: (a) a PNA (peptide nucleic acid) including a polyamide backbone bearing a plurality of ligands at respective spaced locations along the backbone, where each ligand being bound directly or indirectly to a nitrogen atom in the backbone and the ligand bearing nitrogen atoms mainly being separated from one another in the backbone by 4-8 intervening atoms; (b) a nucleic acid analog capable of hybridizing to a nucleic acid of complementary sequence to form a hybrid which is more stable against denaturation by heat than a hybrid between the conventional deoxyribonucleotide and the nucleic acid; or (c) a nucleic acid analogue capable of hybridizing to double-stranded nucleic acid in which one strand has a sequence complementary to the analogue, so as to displace ...

PNA DIAGNOSTIC USE

The present invention pertains to certain nucleic acid analogs and related kits that are useful for the capture, recognition, detection, identification, or quantification of certain chemical or biological entities. 1. A nucleic acid analogue for use in the capture , recognition , detection , identification or quantitation of one or more chemical or microbiological entities , which analogue is(a) a peptide nucleic acid (PNA) comprising a polyamide backbone bearing a plurality of ligands at respective spaced locations along said backbone, said ligands being each independently naturally occurring nucleobases, non-naturally occurring nucleobases or nucleobase-binding groups, each said ligand being bound directly or indirectly to a nitrogen atom in said backbone, and said ligand bearing nitrogen atoms mainly being separated from one another in said backbone by from 4 to 8 intervening atoms;(b) a nucleic acid analogue capable of hybridising to a nucleic acid of complementary sequence to form a hybrid which is more stable against denaturation by heat than a hybrid between the conventional deoxyribonucleotide corresponding to said analogue and said nucleic acid; or(c) a nucleic acid analogue capable of hybridising to a double stranded nucleic acid in which one strand has a sequence complementary to said analogue, so as to displace the other strand from said one strand.54. A nucleic acid analogue as claimed in any one of to claims 1 , incorporating or conjugated to a detectable label.6. A labelled nucleic acid analogue as claimed in claim 5 , wherein said label is a radio isotope label claim 5 , an enzyme label claim 5 , biotin claim 5 , a fluorophore claim 5 , a chemiluminescence label claim 5 , an antigen claim 5 , an antibody or a spin label.7. The use of nucleic acid analogue as defined in any one of to in the capture claim 5 , recognition claim 5 , detection claim 5 , identification or quantitation of one or more chemical or microbiological entities.86. A method of ...

Methods for Determining Sequence Variants Using Ultra-Deep Sequencing

The claimed invention provides for new sample preparation methods enabling direct sequencing of PCR products using pyrophosphate sequencing techniques. The PCR products may be specific regions of a genome. The techniques provided in this disclosure allows for SNP (single nucleotide polymorphism) detection, classification, and assessment of individual allelic polymorphisms in one individual or a population of individuals. The results may be used for diagnostic and treatment of patients as well as assessment of viral and bacterial population identification.

DETECTION AND/OR QUANTITATION OF ENDOTOXIN

Detection and/or quantitation of endotoxin using biolayer interferometry is disclosed. 1. A method of assaying a sample for endotoxin , the method comprising:(a) placing a sample to be assayed in contact with LAL reagent, wherein endotoxin, if present in the sample, reacts with the reagent and a clotted product is formed;(b) contacting a tip of an optical fiber with the sample, wherein clotted product, if present, forms on and/or binds to the tip of the optical fiber, wherein the tip of the optical fiber has an optical thickness, and the optical thickness increases from an initial thickness when clotted product forms on and/or binds to the tip of the optical fiber;(c) determining the optical thickness at the tip of the optical fiber after contact with the sample, wherein an increase in optical thickness from the initial thickness indicates the presence of endotoxin, and an absence of an increase in optical thickness indicates the absence of endotoxin.2. The method of claim 1 , comprising determining a concentration of endotoxin in the sample.3. The method of claim 1 , comprising placing the sample in a sample receiving chamber claim 1 , passing the sample from the sample receiving chamber to a reaction chamber having the LAL claim 1 , reagent therein claim 1 , wherein the tip of the optical fiber contact the sample in the reaction chamber claim 1 , and determining the optical thickness at the tip of the optical fiber after contact with the sample.4. The method of claim 1 , including contacting the tips of a plurality of optical fibers with the sample claim 1 , and determining the optical thicknesses at the tips of the optical fibers after contact with the sample.5. The method of claim 1 , wherein a plurality of optical tips are arranged in a bundle claim 1 , and the method includes contacting the tips of the plurality of optical fibers with the sample claim 1 , and determining the optical thicknesses at the tips of the optical fibers after contact with the sample.6. ...

Asynchronous primed PCR

An asynchronous thermal cycling protocol for nucleic acid amplification uses two primers with thermal melting temperatures different by about 10 to 30° C. After the higher melting primer has annealed and polymerase mediated extension, the uncopied, single-stranded target sequence may be hybridized and detected by a probe. DNA probes may be cleaved by the exonuclease activity of a polymerase. The probe may be a non-cleaving analog such as PNA. When a probe is labelled with a reporter dye and a quencher selected to undergo energy transfer, e.g. FRET, fluorescence from the reporter dye may be effectively quenched when the probe is unbound. Upon hybridization of the probe to complementary target or upon cleavage while bound to target, the reporter dye is no longer quenched, resulting in a detectable amount of fluorescence. The second, lower-melting primer may be annealed and extended to generate a double-stranded nucleic acid. Amplification may be monitored in real time, including each cycle, or at the end point. The asynchronous PCR thermal cycling protocol can generate a preponderance of the PCR amplicon in single-stranded form by repetition at the end of the protocol of annealing and extension of the higher melting primer.

OLIGONUCLEOTIDE ANALOGUE DESCRIBED BY PEN, MONOMERIC SYNTHONES AND PROCEDURES FOR PREPARING THEREOF, AND APPLICATIONS THEREOF

Peptide nucleic acid based molecular sensors for nucleic acids

A chemical moiety including a polymer (P), a first tethering element (T), a ligand (L) which is a specific sequence of PNA, a second tethering element (T′) and a quencher (Q) is disclosed. In the absence of a complement to the PNA sequence, the PNA is in a tightly coiled configuration, thereby quenching the polymer due to the close proximity of the quencher to the polymer. When a receptor is added that recognizes the PNA sequence, a hybridization of the PNA sequence separates the polymer and the quencher, resulting in an increase of detected fluorescence. The same chemistry is advantageously employed in a competitive assay. A method for detecting nucleic acids in a target sample using the PTLT′Q molecule is also disclosed.

Asynchronous primed PCR

An asynchronous thermal cycling protocol for nucleic acid amplification uses two primers with thermal melting temperatures different by about 10 to 30° C. After the higher melting primer has annealed and polymerase mediated extension, the uncopied, single-stranded target sequence may be hybridized and detected by a probe. DNA probes may be cleaved by the exonuclease activity of a polymerase. The probe may be a non-cleaving analog such as PNA. When a probe is labelled with a reporter dye and a quencher selected to undergo energy transfer, e.g. FRET, fluorescence from the reporter dye may be effectively quenched when the probe is unbound. Upon hybridization of the probe to complementary target or upon cleavage while bound to target, the reporter dye is no longer quenched, resulting in a detectable amount of fluorescence. The second, lower-melting primer may be annealed and extended to generate a double-stranded nucleic acid. Amplification may be monitored in real time, including each cycle, or at the end point. The asynchronous PCR thermal cycling protocol can generate a preponderance of the PCR amplicon in single-stranded form by repetition at the end of the protocol of annealing and extension of the higher melting primer.

Asynchronous primed pcr

Asynchronous primed pcr

An asynchronous thermal cycling protocol for nucleic acid amplification uses two primers with thermal melting temperatures different by about 10 to 30 °C. After the higher melting primer has annealed and polymerase mediated extension, the uncopied, single-stranded target sequence may be hybridized and detected by a probe. DNA probes may be cleaved by the exonuclease activity of a polymerase. The probe may be a non-cleaving analog such as PNA. When a probe is labelled with a reporter dye and a quencher selected to undergo energy transfer, e.g. FRET, fluorescence from the reporter dye may be effectively quenched when the probe is unbound. Upon hybridization of the probe to complementary target or upon cleavage while bound to target, the reporter dye is no longer quenched, resulting in a detectable amount of fluorescence. The second, lower-melting primer may be annealed and extended to generate a double-stranded nucleic acid. Amplification may be monitored in real time, including each cycle, or at the end point. The asynchronous PCR thermal cycling protocol can generate a preponderance of the PCR amplicon in single-stranded form by repetition at the end of the protocol of annealing and extension of the higher melting primer.

Asynchronous primed pcr

An asynchronous thermal cycling protocol for nucleic acid amplification uses two primers with thermal melting temperatures different by about 10 to 30 ~C . After the higher melting primer has annealed and polymerase mediated extension, the uncopied, single-stranded target sequence may be hybridized a nd detected by a probe. DNA probes may be cleaved by the exonuclease activity o f a polymerase. The probe may be a non-cleaving analog such as PNA. When a pro be is labelled with a reporter dye and a quencher selected to undergo energy transfer, e.g. FRET, fluorescence from the reporter dye may be effectively quenched when the probe is unbound. Upon hybridization of the probe to complementary target or upon cleavage while bound to target, the reporter dy e is no longer quenched, resulting in a detectable amount of fluorescence. The second, lower-melting primer may be annealed and extended to generate a doub le- stranded nucleic acid. Amplification may be monitored in real time, includin g each cycle, or at the end point. The asynchronous PCR thermal cycling protoc ol can generate a preponderance of the PCR amplicon in single-stranded form by repetition at the end of the protocol of annealing and extension of the high er melting primer.

Asynchronous primed PCR

An asynchronous thermal cycling protocol for nucleic acid amplification uses two primers with thermal melting temperatures different by about 10 to 30 °C. After the higher melting primer has annealed and polymerase mediated extension, the uncopied, single-stranded target sequence may be hybridized and detected by a probe. DNA probes may be cleaved by the exonuclease activity of a polymerase. The probe may be a non-cleaving analog such as PNA. When a probe is labeled with a reporter dye and a quencher selected to undergo energy transfer, e.g. FRET, fluorescence from the reporter dye may be effectively quenched when the probe is unbound. Upon hybridization of the probe to complementary target or upon cleavage while bound to target, the reporter dye is no longer quenched, resulting in a detectable amount of fluorescence. The second, lower-melting primer may be annealed and extended to generate a double-stranded nucleic acid. Amplification may be monitored in real time, including each cycle, or at the end point. The asynchronous PCR thermal cycling protocol can generate a preponderance of the PCR amplicon in single-stranded form by repetition at the end of the protocol of annealing and extension of the higher melting primer.

Asynchronous primed PCR

An asynchronous thermal cycling protocol for nucleic acid amplification uses two primers with thermal melting temperatures different by about 10 to 30 °C. After the higher melting primer has annealed and polymerase mediated extension, the uncopied, single-stranded target sequence may be hybridized and detected by a probe. DNA probes may be cleaved by the exonuclease activity of a polymerase. The probe may be a non-cleaving analog such as PNA. When a probe is labeled with a reporter dye and a quencher selected to undergo energy transfer, e.g. FRET, fluorescence from the reporter dye may be effectively quenched when the probe is unbound. Upon hybridization of the probe to complementary target or upon cleavage while bound to target, the reporter dye is no longer quenched, resulting in a detectable amount of fluorescence. The second, lower-melting primer may be annealed and extended to generate a double-stranded nucleic acid. Amplification may be monitored in real time, including each cycle, or at the end point. The asynchronous PCR thermal cycling protocol can generate a preponderance of the PCR amplicon in single-stranded form by repetition at the end of the protocol of annealing and extension of the higher melting primer.

Asynchronous primed PCR

An asynchronous thermal cycling protocol for nucleic acid amplification uses two primers with thermal melting temperatures different by about 10 to 30° C. After the higher melting primer has annealed and polymerase mediated extension, the uncopied, single-stranded target sequence may be hybridized and detected by a probe. DNA probes may be cleaved by the exonuclease activity of a polymerase. The probe may be a non-cleaving analog such as PNA. When a probe is labelled with a reporter dye and a quencher selected to undergo energy transfer, e.g. FRET, fluorescence from the reporter dye may be effectively quenched when the probe is unbound. Upon hybridization of the probe to complementary target or upon cleavage while bound to target, the reporter dye is no longer quenched, resulting in a detectable amount of fluorescence. The second, lower-melting primer may be annealed and extended to generate a double-stranded nucleic acid. Amplification may be monitored in real time, including each cycle, or at the end point. The asynchronous PCR thermal cycling protocol can generate a preponderance of the PCR amplicon in single-stranded form by repetition at the end of the protocol of annealing and extension of the higher melting primer.

Asynchronous primed pcr

An asynchronous thermal cycling protocol for nucleic acid amplification uses two primers with thermal melting temperatures different by about 10 to 30 °C. After the higher melting primer has annealed and polymerase mediated extension, the uncopied, single-stranded target sequence may be hybridized and detected by a probe. DNA probes may be cleaved by the exonuclease activity of a polymerase. The probe may be a non-cleaving analog such as PNA. When a probe is labelled with a reporter dye and a quencher selected to undergo energy transfer, e.g. FRET, fluorescence from the reporter dye may be effectively quenched when the probe is unbound. Upon hybridization of the probe to complementary target or upon cleavage while bound to target, the reporter dye is no longer quenched, resulting in a detectable amount of fluorescence. The second, lower-melting primer may be annealed and extended to generate a double-stranded nucleic acid. Amplification may be monitored in real time, including each cycle, or at the end point. The asynchronous PCR thermal cycling protocol can generate a preponderance of the PCR amplicon in single-stranded form by repetition at the end of the protocol of annealing and extension of the higher melting primer.

Asynchronous primed pcr

An asynchronous thermal cycling protocol for nucleic acid amplification uses two primers with thermal melting temperatures different by about 10 to 30 °C. After the higher melting primer has annealed and polymerase mediated extension, the uncopied, single-stranded target sequence may be hybridized and detected by a probe. DNA probes may be cleaved by the exonuclease activity of a polymerase. The probe may be a non-cleaving analog such as PNA. When a probe is labelled with a reporter dye and a quencher selected to undergo energy transfer, e.g. FRET, fluorescence from the reporter dye may be effectively quenched when the probe is unbound. Upon hybridization of the probe to complementary target or upon cleavage while bound to target, the reporter dye is no longer quenched, resulting in a detectable amount of fluorescence. The second, lower-melting primer may be annealed and extended to generate a double-stranded nucleic acid. Amplification may be monitored in real time, including each cycle, or at the end point. The asynchronous PCR thermal cycling protocol can generate a preponderance of the PCR amplicon in single-stranded form by repetition at the end of the protocol of annealing and extension of the higher melting primer.

Asynchronous primed pcr

An asynchronous thermal cycling protocol for nucleic acid amplification uses two primers with thermal melting temperatures different by about 10 to 30 °C. After the higher melting primer has annealed and polymerase mediated extension, the uncopied, single-stranded target sequence may be hybridized and detected by a probe. DNA probes may be cleaved by the exonuclease activity of a polymerase. The probe may be a non-cleaving analog such as PNA. When a probe is labelled with a reporter dye and a quencher selected to undergo energy transfer, e.g. FRET, fluorescence from the reporter dye may be effectively quenched when the probe is unbound. Upon hybridization of the probe to complementary target or upon cleavage while bound to target, the reporter dye is no longer quenched, resulting in a detectable amount of fluorescence. The second, lower-melting primer may be annealed and extended to generate a double-stranded nucleic acid. Amplification may be monitored in real time, including each cycle, or at the end point. The asynchronous PCR thermal cycling protocol can generate a preponderance of the PCR amplicon in single-stranded form by repetition at the end of the protocol of annealing and extension of the higher melting primer.

Multipartite high-affinity nucleic acid probes

The invention provides a collection of probes useful for hybridizing to a target nucleic acid. The probes associate with each other, binding with high affinity to the target nucleic acid, to form three-way junctions and other complexes. At least one of the probes in each collection includes a nucleic acid analog. Methods using the probes in hybridization and as primers are also provided.

Peptide nucleic acids having amino acid side chains

A novel class of compounds, known as peptide nucleic acids, bind complementary DNA and RNA strands more strongly than the corresponding DNA or RNA strands, and exhibit increased sequence specificity and solubility. The peptide nucleic acids comprise ligands selected from a group consisting of naturally-occurring nucleobases and non-naturally-occurring nucleobases attached to a polyamide backbone, and contain alkylamine side chains.

Template-dependent ligation with PNA-DNA chimeric probes

The invention provides methods, kits, and compositions for ligation of PNA-DNA chimeric probes and oligonucleotides when they are hybridized adjacently to template nucleic acids using ligases and ligation reagents. Structural requirements of the chimeras for ligation include 5 to 15 contiguous PNA monomer units, 2 or more contiguous nucleotides, and a 3′ hydroxyl or 5′ hydroxyl terminus. The chimera and/or oligonucleotide may be labelled with fluorescent dyes or other labels. The methods include, for example, oligonucleotide-ligation assays (OLA) and single nucleotide polymorphism detection.

Peptide nucleic acids

A novel class of compounds, known as peptide nucleic acids, bind complementary ssDNA and RNA strands more strongly than a corresponding DNA. The peptide nucleic acids generally comprise ligands such as naturally occurring DNA bases attached to a peptide backbone through a suitable linker.

Peptide nucleic acids having enhanced binding affinity, sequence specificity and solubility

A novel class of compounds, known as peptide nucleic acids, bind complementary DNA and RNA strands more strongly than a corresponding DNA strand, and exhibit increased sequence specificity and solubility. The peptide nucleic acids comprise ligands selected from a group consisting of naturally-occurring nucleobases and non-naturally-occurring nucleobases attached to a polyamide backbone, and contain C 1 -C 8 alkylamine side chains. Methods of enhancing the solubility, binding affinity and sequence specificity of PNAs are provided.

Peptide nucleic acids having enhanced binding affinity and sequence specificity

A novel class of compounds, known as peptide nucleic acids, bind complementary DNA and RNA strands more strongly than a corresponding DNA strand, and exhibit increased sequence specificity and binding affinity. Methods of increasing binding affinity and sequence specificity of peptide nucleic acids are provided wherein some peptide nucleic acids comprise ligands selected from a group consisting of naturally-occurring nucleobases and non-naturally-occurring nucleobases attached to a polyamide backbone, while other peptide nucleic acids contain at least one 2,6-diaminopurine nucleobase and at least one C1-C8 alkylamine side chain.

Peptide nucleic acids

A novel class of compounds, known as peptide nucleic acids, bind complementary ssDNA and RNA strands more strongly than a corresponding DNA. The peptide nucleic acids generally comprise ligands such as naturally occurring DNA bases attached to a peptide backbone through a suitable linker.

Nucleic acid amplification

Disclosed are compositions and a method for amplification of nucleic acid sequences of interest. The method is based on strand displacement replication of the nucleic acid sequences by primers. The disclosed method is a form of multiple displacement amplification (MDA) useful for amplifying genomic nucleic acid samples and other nucleic acid samples of high complexity. The disclosed method can be used to amplify such highly complex nucleic acid samples using only one or a limited number of primers. It has been discovered that one or a small number of primers can effectively amplify whole genomes an other nucleic acid samples of high sequence complexity. The primers are specially selected or designed to be able to prime and efficiently amplify the broad range of sequences present in highly complex nucleic acid samples despite the limited amount of primer sequence represented in the primers. It has been discovered that generation of high molecular weight artifacts, in an isothermal amplification procedure, is substantially reduced or eliminated while still allowing the desired amplification of input DNA by carrying out the reaction at a higher temperature and, optionally, in the presence of one or more additives. It has also bee discovered that amplification reactions can produce amplification products of high quality, such as low amplification bias, if performed on an amount of nucleic acid at or over a threshold amount and/or on nucleic acids at or below a threshold concentration.

Uses of nucleic acid analogues in the inhibition on nucleic acid amplification

Nucleic acid analogues of the kind described in PCT/EP92/01220 (PNA's) which hybridise strongly to nucleic acids are used to inhibit nucleic acid amplification procedures such as PCR. False positives in subsequent PCR assays are prevented by hybridising a PNA to PCR amplification products. Assays capable of discriminating between single base pair mutants are conducted by using a PNA hybridising to one of the two allelic forms to inhibit a PCR amplification of that form selectively. Asymmetric PCR amplifications are carried out by starting a PCR symmetricaly using like quantities of forward and reverse primers and once the amplification is established disabling one primer by hybridising a PNA thereto.

Process for the preparation of 6-[[(alkoxy)carbonyl]amino]-9H-Purine-9-acetic acid and 4-[(alkoxycarbonyl)amino]-2-oxo-1(2H)-Pyrimidineacetic acid derivatives as synthons for the synthesis of peptide nucleic acids

A process is disclosed for the preparation of 6-[[(alkoxy)carbonyl]amino]-9H-Purine-9-acetic acid and 4-[(alkoxycarbonyl)amino]-2-oxo-1(2H)-Pyrimidineacetic acid derivatives as synthons for the synthesis of peptide nucleic acids: or

Peptide nucleic acids

A novel class of compounds, known as peptide nucleic acids, bind complementary ssDNA and RNA strands more strongly than a corresponding DNA. The peptide nucleic acids generally comprise ligands such as naturally occurring DNA bases attached to a peptide backbone through a suitable linker.

Strand displacement methods employing competitor oligonucleotides for isolating one strand of a double-stranded nucleic acid

The present invention provides methods and kits for isolating one strand of a double-stranded target nucleic acid. The method capitalizes on the differences in the kinetics and thermodynamic stabilities between conventional DNA/DNA, DNA/RNA and RNA/RNA duplexes and heteroduplexes in which one strand of the heteroduplexe is a nucleobase polymer having a net positively charged or net neutral backbone, such as a PNA.

Use of nucleic acid analogues in inhibiting nucleic acid amplification

Compositions of solvents and high concentrations of nucleic acid analogs

The invention provides compositions and methods suitable for dissolving nucleic acid analogs with uncharged, neutral backbones at high concentration s at approximately neutral pH. By using the compositions of the invention, whi ch include a polar, aprotic solvent, concentrations of nucleic acid analogs suc h as PNA can be achieved in the range of approximately 1 .mu.M to 10 mM.</SDOA B>

Peptide nucleic acids having 2,6-diaminopurine nucleobases

A novel class of compounds, known as peptide nucleic acids, bind complementary DNA and RNA strands more strongly than a corresponding DNA strand, and exhibit increased sequence specificity and binding affinity. The peptide nucleic acids of the invention comprise ligands selected from a group consisting of naturally-occurring nucleobases and non-naturally-occurring nucleobases attached to a polyamide backbone. Some PNAs of the invention also contain C 1 -C 8 alkylamine side chains.

The use of nucleic acid analogues in diagnostics and analytical procedures.

Procédés de capture, de reconnaissance, de détection, d'identification ou de quantification d'acides nucléiques et utilisations à des fins de diagnostic, dans lesquels sont utilisés (a) un acide nucléique peptidique (PNA) comprenant un squelette de polyamide portant une pluralité de ligands en des emplacements espacés respectifs le long dudit squelette, lesdits ligands étant chacun des nucléobases se produisant naturellement indépendamment, des nucléobases non-naturelles ou des groupes de liaison de nucléobases, chacun desdits ligands étant lié directement ou indirectement à un atome d'azote dans ledit squelette, et lesdits atomes d'azote porteurs de ligands étant essentiellement séparés les uns des autres dans ledit squelette par quatre à huit atomes de séparation; (b) un analogue d'acide nucléique pouvant s'hybrider avec un acide nucléique d'une séquence complémentaire afin de former un hybride plus stable contre la dénaturation sous l'effet de la chaleur qu'un hybride se trouvant entre le désoxyribonucléotide classique correspondant audit analogue et ledit acide nucléique; ou (c) un analogue d'acide nucléique capable de s'hybrider avec un acide nucléique bicaténaire dans lequel un brin a une séquence complémentaire audit analogue, de manière à déplacer l'autre brin dudit brin. Les composés préférés ont la formule (III), dans laquelle chaque L est choisi indépendament dans le groupe constitué par hydrogène, phényle, nucléobases naturelles, et nucléobases non naturelles; chaque R7' est choisi indépendament dans le groupe constitué par hydrogène et les chaînes latérales d'acides aminés alpha naturelles; l représente un nombre entier compris entre 1 et 60, k et m représentent chacun indépendament 0 ou 1; et chaque l est compris indépendament entre 0 et 5; Rh représente OH, NH2 ou NHLysNH2; et Ri représente H ou COCH3. Methods for capture, recognition, detection, identification or quantification of nucleic acids ...

Double-stranded peptide nucleic acids

A novel class of compounds, known as peptide nucleic acids, form double-stranded structures with one another and with ssDNA. The peptide nucleic acids generally comprise ligands such as naturally occurring DNA bases attached to a peptide backbone through a suitable linker.

Nucleic acid analogue induced transcription of double stranded dna

RNA is transcribed from a double stranded DNA template by forming a complex by hybridising to the template at a desired transcription initiation site one or more oligonucleic acid analogues of the PNA type capable of forming a transcription initiation site with the DNA and exposing the complex to the action of a DNA dependant RNA polymerase in the presence of nucleoside triphosphates. Equal length transcripts may be obtained by placing a block to transcription downstream from the initiation site or by cutting the template at such a selected location. The initiation site is formed by displacement of one strand of the DNA locally by the PNA hybridisation.

Peptide nucleic acids

Asynchronous primed PCR

Use of nucleic acid analogues in the inhibition of nucleic acid amplification

Nucleic acid analogues of the kind described in PCT/EP92/01220 (PNA's) which hybridise strongly to nucleic acids are used to inhibit nucleic acid amplification procedures such as PCR. False positives in subsequent PCR assays are prevented by hybridising a PNA to PCR amplification products. Assays capable of discriminating between single base pair mutants are conducted by using a PNA hybridising to one of the two allelic forms to inhibit a PCR amplification of that form selectively. Asymmetric PCR amplifications are carried out by starting a PCR symmetricaly using like quantities of forward and reverse primers and once the amplification is established disabling one primer by hybridising a PNA thereto.

Process for the inhibition of nucleic acid amplification using nucleic acid analogues

A találmány tárgyát nukleinsav-amplifikációs eljárások, mint például aPCR gátlása képezi nukleinsavanalógok (PNA-k) segítségével, amelyekerősen hibridizálódnak a nukleinsavakhoz. A további PCR eljárásokban ahamis pozitív eredmények létrejöttének megakadályozására egy PNA-t aPCR nukleinsav-amplifikációs termékhez hibridizálnak. Az egy bázispármutációját felismerő vizsgálatokat úgy hajtják végre, hogy egy olyanPNA-t használnak, amely a két allél forma közül az egyikhezszelektíven hibridizálódik, és ezáltal szelektíven megakadályozzaannak a formának a PCR amplifikációját. Aszimmetrikus PCRamplifikációkat úgy végeznek, hogy egy PCR reakciót szimmetrikusanindítanak el, az előrevivő és a fordított indítómolekulákból hasonlómennyiséget használva, majd miután létrejött az amplifikáció, az egyikindítómolekulát egy PNA hozzáhibridizálásával működésképtelennéteszik. ŕ

Improved synthons for the synthesis and deprotection of peptide nucleic acids under mild conditions

Peptide nucleic acid conjugates

Peptide nucleic acids complexes of two peptide nucleic acid strands and one nucleic acid strand

Peptide nucleic acids and analogues of peptide nucleic acids are used to form duplex, triplex, and other structures with nucleic acids and to modify nucleic acids. The peptide nucleic acids and analogues thereof also are used to modulate protein activity through, for example, transcription arrest, transcription initiation, and site specific cleavage of nucleic acids.

Methods for determining sequence variants using ultra-deep sequencing

The claimed invention provides for new sample preparation methods enabling direct sequencing of PCR products using pyrophosphate sequencing techniques. The PCR products may be specific regions of a genome. The techniques provided in this disclosure allows for SNP (single nucleotide polymorphism) detection, classification, and assessment of individual allelic polymorphisms in one individual or a population of individuals. The results may be used for diagnostic and treatment of patients as well as assessment of viral and bacterial population identification.

Template-dependent ligation with pna-dna chimeric probes

Polymerase extension at 3' terminus of pna-dna chimera

The invention provides methods and a kit for primer extension of PNA-DNA chimera from template nucleic acids using polymerases, nucleotide 5'- triphosphates, and primer extension reagents. Structural requirements of the chimera for primer extension include 5 to 15 contiguous PNA monomer units, 3 or more contiguous nucleotides, and a 3' hydroxyl terminus. The chimera and/ or a nucleotide is labelled with the fluorescent dyes or other labels. The methods include DNA sequencing, DNA fragment analysis, reverse transcription , mini-sequencing, chromosome labelling, amplification, and single nucleotide polymorphism (SNP) detection.

Methods for isolating one strand of a double-stranded nucleic acid

The present invention provides methods and kits for isolating one strand of a double-stranded target nucleic acid. The method capitalizes on the differences in the kinetics and thermodynamic stabilities between conventional DNA/DNA, DNA/RNA and RNA/RNA duplexes and heteroduplexes in which one strand of the heteroduplexe is a nucleobase polymer having a net positively charged or net neutral backbone, such as a PNA.

Methods for determining sequence variants using ultra-deep sequencing

The claimed invention provides for new sample preparation methods enabling direct sequencing of PCR products using pyrophosphate sequencing techniques. The PCR products may be specific regions of a genome. The techniques provided in this disclosure allows for SNP (single nucleotide polymorphism) detection, classification, and assessment of individual allelic polymorphisms in one individual or a population of individuals. The results may be used for diagnostic and treatment of patients as well as assessment of viral and bacterial population identification.

Linked peptide nucleic acids

Novel peptide nucleic acids are disclosed which comprise nucleobases including C-pyrimidines and iso-pyrimidines. Suitable C-pyrimidines include pseudo-isocytosine and pseudo-uracil. Suitable iso-pyrimidine bases include iso-cytosine. Such compositions typically exhibit increased binding affinity.

Peptide nucleic acids

Methods for determining sequence variants using ultra-deep sequencing

The claimed invention provides for new sample preparation methods enabling direct sequencing of PCR products using pyrophosphate sequencing techniques. The PCR products may be specific regions of a genome. The techniques provided in this disclosure allows for SNP (single nucleotide polymorphism) detection, classification, and assessment of individual allelic polymorphisms in one individual or a population of individuals. The results may be used for diagnostic and treatment of patients as well as assessment of viral and bacterial population identification.

Peptide nucleic acids

A novel class of compounds, known as peptide nucleic acids, bind complementary ssDNA and RNA strands more strongly than a corresponding DNA. The peptide nucleic acids generally comprise ligands such as naturally occurring DNA bases attached to a peptide backbone through a suitable linker.

Nucleic acid analogue, its use, in vitro method and kit

Paired end sequencing

The present invention provides for a method of preparing a target nucleic acid fragments to produce a smaller nucleic acid which comprises the two ends of the target nucleic acid. Specifically, the invention provides cloning and DNA manipulation strategies to isolate the two ends of a large target nucleic acid into a single small DNA construct for rapid cloning, sequencing, or amplification.

Methods for isolating one strand of a double-stranded nucleic acid

The present invention provides methods and kits for isolating one strand of a double-stranded target nucleic acid. The method capitalizes on the differences in the kinetics and thermodynamic stabilities between conventional DNA/DNA, DNA/RNA and RNA/RNA duplexes and heteroduplexes in which one strand of the heteroduplexe is a nucleobase polymer having a net positively charged or net neutral backbone, such as a PNA.

Binary probe and clamp composition and methods for target hybridization detection

Binary probe and clamp compositions conduct methods for target hybridization detection. Where the probe is a substrate for exonuclease cleavage, the composition provides quantitation and detection of PCR products, by real-time and end-point measurements. Where the probe is an amplification primer, the composition provides an improved method for labelling and detection of PCR products. Probes and clamps may be labelled with fluorescent dyes, quenchers, hybridization-stabilizing moieties, chemiluminescent dyes, and affinity ligands. Clamps may be nucleic acid analogs, such as 2-aminoethylglycine PNA.

Inhibition method of nucleic acid amplification, and a set for nucleic acid amplification

PCT No. PCT/EP93/01435 Sec. 371 Date Mar. 10, 1995 Sec. 102(e) Date Mar. 10, 1995 PCT Filed Jun. 7, 1993 PCT Pub. No. WO93/25706 PCT Pub. Date Dec. 23, 1993Nucleic acid analogues such as peptide-nucleic acids (PNAs) which hybridize strongly to nucleic acids are used to inhibit nucleic acid amplification procedures such as PCR. False positives in subsequent PCR assays are prevented by hybridizing a PNA to PCR amplification products. Assays capable of discriminating between single base mutants are conducted by using a PNA hybridizing to one of the two allelic forms to inhibit a PCR amplification of that form selectively. Asymmetric PCR amplifications are carried out by starting a PCR symmetrically using like quantities of forward and reverse primers, and, once the amplification is established, disabling one primer by hybridizing a PNA thereto.

Paired end sequencing

The present invention provides for a method of preparing a target nucleic acid fragments to produce a smaller nucleic acid which comprises the two ends of the target nucleic acid. Specifically, the invention provides cloning and DNA manipulation strategies to isolate the two ends of a large target nucleic acid into a single small DNA construct for rapid cloning, sequencing, or amplification.

Binärsonde, klammerzusammensetzung und verfahren zum zielhybridisierungsnachweis

Peptid-nukleinsäure-konjugate

Verlängerung einer pna-dna chimäre an ihrem 3' ende mittels einer polymerase

Asynchrones pcr priming

Mittels nukleinsäureanalogs induzierte transkription von doppelsträngiger dna

Zusammensetzungen von lösungsmitteln und hochkonzentrierte nukleinsäureanaloga

Struktur höherer ordnung und bindung von peptidnukleinsäuren

エンドトキシンの検出及び／又は定量

【課題】エンドトキシンを対象として試料をアッセイする方法を提供する。【解決手段】（ａ）アッセイの対象となる試料をＬＡＬ試薬と接触させ、エンドトキシンが上記試料中に存在する場合にはエンドトキシンが前記試薬と反応して凝固生成物が形成されるステップと、（ｂ）光ファイバーの端部を前記試料と接触させ、凝固生成物が存在する場合には凝固生成物が上記光ファイバーの上記端部上に形成する及び／又は上記端部に結合し、上記光ファイバーの上記端部がある光学的厚さを有し、凝固生成物が上記光ファイバーの上記端部上に形成する及び／又は上記端部に結合すると、上記光学的厚さが当初の厚さから増加するステップと、（ｃ）上記試料との接触後、上記光ファイバーの上記端部における上記光学的厚さを測定するステップとを含む、方法。【選択図】なし

Doppelsträngige peptidenukleinsäuren

Template abhängige ligierung mit pna-dna chimerischen sonden

Peptidnukleinsäuren mit erhöhter bindungsaffinität, sequenzspezifität und löslichkeit

Verwendung von nuklein-säure analoge in der inhibition von nuklein-säure amplifikation

平板スクリーンを洗浄するために方向付けられたハイドロバーストシステム

【課題】複雑なトラフタイプのマニホールドを作成せずに平板型スクリーンを効果的に洗浄できるようにする。 【解決手段】クリーン取水システム１００は、開口端と室とを持つボディを具え、平板スクリーン８０は、ボディの開口端に配置される。空気バーストシステム１００は、室内に配置された複数の管を具え、平板スクリーン８０からゴミを取り除く。管は、平板スクリーン８０に近接して互いに平行に配置され、各管は平板スクリーン８０に面する側に複数のオリフィスを具え、且つ、複数の管の後ろ側に沿って配置され、ディレクタのそれぞれはチャンネルを有し、そのチャンネルに管が配置される。圧縮空気がタンクからバルブにより放出され、空気バーストが管のオリフィスから散布される。ディレクタのそれぞれは、散布の結果として得られるオリフィスからの水／空気バーストを近接する平板スクリーン８０に向けて、平板スクリーン８０からゴミを取り除く。 【選択図】図４Ｂ

Verknüpfte peptid-nukleinsäuren

両末端配列決定（ｐａｉｒｅｄｅｎｄｓｅｑｕｅｎｃｉｎｇ）

【課題】両末端配列決定アプローチおよび他の核酸技術に有用な新規の方法、システムおよび組成物を提供する。 【解決手段】本発明は、核酸の大きいフラグメントの両方の末端を単離および配列決定するための迅速でありかつ費用対効果の高い方法を提供し、この方法は、迅速であり、かつ自動化に適しており、そしてＤＮＡの大きいフラグメントの配列決定および連鎖を可能にする。本発明の方法は、ＤＮＡフラグメントのライブラリーを産生するために使用され得、そのフラグメントは、ＤＮＡのより大きいフラグメント由来の末端を含む。 【選択図】なし

Peptidnucleinsäuren

Peptidnukleinsäuren