文献类型：专利

英文题名：Quantitative detection of RNA polymerase and ligase mediated coupling reactions for PCR analysis of nucleic acids, comprises using a target nucleic acid and a oligonucleotide nucleotide probe for hybridization

作者：YIN B;YE B;YU C

机构：[1]UNIV EAST CHINA SCI & TECHNOLOGY

申请号：CN104032031-A

申请日：2014-07-04

公开日：2014-09-10

语种：英文

收录：DERWENT

摘要：NOVELTY - Quantitative detection of RNA polymerase and ligase mediated coupling reactions for PCR analysis of nucleic acids, comprises: using a target nucleic acid and a oligonucleotide nucleotide probe for hybridization, connection cut-ligase catalyzed oligonucleotide probe to obtain a connection between the product of the ligation product sequence containing the RNA polymerase promoter; RNA polymerase promoter sequence of the forward primer containing a hybridize with the ligated product to form a double-stranded promoter transcription template structure. USE - The method is useful for the quantitative detection of RNA polymerase and ligase mediated coupling reactions for PCR analysis of nucleic acids (claimed). ADVANTAGE - The method has high sensitivity, strong specificity, simple, short and can be used in clinical diagnosis. DETAILED DESCRIPTION - Quantitative detection of RNA polymerase and ligase mediated coupling reactions for PCR analysis of nucleic acids, comprises: using a target nucleic acid and a oligonucleotide nucleotide probe for hybridization, connection cut-ligase catalyzed oligonucleotide probe to obtain a connection between the product of the ligation product sequence containing the RNA polymerase promoter; RNA polymerase promoter sequence of the forward primer containing a hybridize with the ligated product to form a double-stranded promoter transcription template structure; RNA polymerase recognition of the transcriptional template to synthesize the promoter and downstream of the RNA fragments complementary to the probe sequence, the RNA can serve as target nucleic acid fragments with oligonucleotide probe hybridization, ligation-mediated connection between the enzyme-catalyzed nucleotide probe oligonucleotide; in PCR reactions, DNA polymerase, the forward primer, reverse primer, and deoxynucleotide triphosphates (dNTPs) under the combined effect thereof, to connect the product as a template to obtain a large amount of double-stranded DNA, SYBR Green I dye binding to double-stranded DNA to generate a fluorescent signal in real time in the reaction system was measured fluorescence signal intensity with the standard contrast curve to calculate the concentration of the target nucleic acid.