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Methods for the detection and quantification of dna by single-molecule hybridization and manipulation

A single-stranded nucleic acid molecule, molecular technology, applied to DNA,] the present invention relates to various applications, detection and quantification for rapid DNA or RNA renaturation of denatured double-stranded nucleic acid, chromosome distribution or gene expression analysis Existence of specific nucleic acid molecules, detection and quantification of nucleic acids, can solve the problems of increased overall cost, time-consuming, easy pollution, etc., achieve rapid and large-scale diagnosis, and improve the effect of signal/noise ratio

Active Publication Date: 2017-04-26
CENT NAT DE LA RECHERCHE SCI +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, all methods developed so far have serious disadvantages
In particular, they all use labeled nucleotides (e.g., using fluorescent labels), thus causing a drastic increase in overall cost
Additionally, amplifying target sequences is time consuming, increases the potential for errors, and is highly susceptible to contamination

Method used

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  • Methods for the detection and quantification of dna by single-molecule hybridization and manipulation
  • Methods for the detection and quantification of dna by single-molecule hybridization and manipulation
  • Methods for the detection and quantification of dna by single-molecule hybridization and manipulation

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Experimental program
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Embodiment Construction

[0147] method

[0148] DNA hairpin construction

[0149] like Figure 4 Short DNA hairpins (in figure 1 and figure 2 83bp in, or in figure 2 middle 179bp). In the first step, the dH 2Oligonucleotide (oligo) A-1 in O (SEQ ID NO. 3: 5'-biotin-TTT TTT TTT TTT TTT TTT TTT TTT TTT TTTTTT TTT TTT TTT TTT TTTTTT TTG GAT TCG CGG GTC TCT-3') and oligonucleotide A-2 (SEQ ID NO.4:5'-AAC CGT CCT TTA CTT GTC ATG CGC TCT AAT CTC TGG GCA TCT GGC TAT GATGTT GAT GGA ACT GAC CAA ACG TCG GTG GG-3') annealed to complementary Oligonucleotide A-3 (SEQ ID NO. 5: 5'-phosphorylated AGG AAG AGA CCC GCG AAT CCC CCA CCG ACG TTT GGT CAG TT-3'). These annealed products, labeled fraction A, were washed using the NucleoSpin Extract II kit (Clontech). For oligonucleotide B-1 corresponding to the middle part of the 83 bp hairpin (SEQ ID NO. 6: 5'-phosphorylated-TCC TGA TTG TCATCG GCT GGC TGT TCG GTT AGT TTC GAA GAC TT-3') and oligonucleotide B-2 (SEQ ID NO.7:5'-phosphorylated-GCG AAA GTC TTC GAA ACT...

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Abstract

The present invention relates to rapid methods for the detection and quantification of nucleic acids, DNA or RNA. In particular, the present invention provides methods for detecting and quantifying the presence of specific nucleic acid molecules based on physical and electronic processes. The methods of the invention are particularly suitable for a variety of applications in the detection of abnormal chromosome distribution or gene expression analysis.

Description

Background technique [0001] The present invention relates to rapid methods for the detection and quantification of nucleic acids, DNA or RNA. In particular, the present invention provides methods for detecting and quantifying the presence of specific nucleic acid molecules based on physical and electronic processes. The methods of the invention are particularly suitable for a variety of applications in the detection of abnormal chromosome distribution or gene expression analysis. [0002] The detection and quantification of nucleic acid sequences is important to a wide range of commonly utilized biological applications. The applications include the field of in vitro diagnostics, including clinical diagnostics, research in the field of molecular biology, high throughput drug screening, veterinary diagnostics, agricultural genetics testing, environmental testing, food testing, industrial engineering monitoring and insurance testing. In particular, demonstrating the presence of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6825C12Q1/6883C12Q1/6886C12Q2600/156C12Q2523/119C12Q2523/305C12Q1/6816C12Q2525/301C12Q1/6813C12Q1/6844
Inventor D·本西蒙J-F·阿莱曼德F-Y·丁V·克罗凯特
Owner CENT NAT DE LA RECHERCHE SCI