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Capture of target DNA and RNA by probes comprising intercalator molecules

An intercalator, target technology, applied in DNA/RNA fragments, recombinant DNA technology, biochemical equipment and methods, etc., can solve problems such as insufficient identification of base mismatches

Inactive Publication Date: 2014-01-15
QUANTIBACT AS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Other limitations of known methods for the diagnosis and detection of short nucleotide sequences are that they usually involve at least one purification step and that they are not specific enough to recognize a single base mismatch

Method used

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  • Capture of target DNA and RNA by probes comprising intercalator molecules
  • Capture of target DNA and RNA by probes comprising intercalator molecules
  • Capture of target DNA and RNA by probes comprising intercalator molecules

Examples

Experimental program
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Effect test

Embodiment

[0517] Assessing the effect of abasic sites on Tm by obtaining melting curves

[0518] use A Fluorescence Resonance Energy Transfer (FRET) system on 2.0 evaluates changes in melting point (Tm) caused by base mismatch or debasic sites (B) in two different oligonucleotide sequences.

[0519] Oligonucleotides were purchased from IBM GmbH ( Germany) or DNA Technology A / S (Risskov, Denmark), high performance liquid chromatography (HPLC) purification and subsequent quality control were performed on a 0.2 μmol synthesis scale. Oligonucleotides were synthesized using 3' amino-modifier-C7 followed by attachment to ATTO495NHS-ester, or using 5' amino-modifier-C6 followed by attachment to ATTO590NHS-ester. ATTO495 acts as a FRET donor and is a modification of acridine orange with an excitation maximum at 495nm and an emission maximum at 527nm. ATTO590 is a derivative of Rhodamine dye with an excitation maximum at 594nm and an emission maximum at 624nm. Pair ATTO495 / ATTO590FRET in ...

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Abstract

The present invention relates to a technology for specific capture of single stranded target Polynucleotide by a complementary probe comprising one or more intercalator molecules. The method further involves removal of one or more types of bases in the single stranded target Polynucleotide prior to interaction with the complementary probe. This results in generation of one or more abasic sites which can interact with and / or into where the intercalator molecule can be inserted.

Description

technical field [0001] The present invention relates to a molecular diagnostic technique involving the specific capture of single-stranded polynucleotides of interest, which may be made from naturally occurring nucleotides, by complementary probes comprising one or more intercalator molecules. or may be made from nucleotides not known in nature, or any mixture thereof, such as DNA and / or RNA. The method further involves removing one or more types of bases from a target polynucleotide, which may be made from naturally occurring nucleotides, prior to the interaction of the target polynucleotide with a complementary probe. or may be made from nucleotides not known in nature, or any mixture thereof, such as DNA and / or RNA. Background technique [0002] Polymerase chain reaction (PCR) is a technique widely used in molecular genetics and diagnostics that can analyze any short DNA sequence, even if the sample contains only low levels of DNA. PCR is used to amplify selected fragme...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6816C12Q1/6827C12Q1/6883C12Q2563/173C12Q2525/119C12Q2521/531C12Q2523/125C12N15/11Y02A50/30
Inventor 乌费·维斯特·施奈德尼娜·约恩克简·G·里斯贝
Owner QUANTIBACT AS
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