RNA one-step-lysis and one-tube RT-PCR detection method

A technology of RT-PCR and detection method, which is applied in the field of RNA one-step cleavage and one-tube RT-PCR detection, which can solve problems such as difficult control of condition optimization, high complexity and difficulty, non-specific amplification and false positives, and achieve Improve reaction efficiency and success rate, reduce non-specific reactions, and reduce the effect of occurrence probability
CN103333955AActive Publication Date: 2013-10-02ZHEJIANG JFK BIOLOGICAL TECH
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
ZHEJIANG JFK BIOLOGICAL TECH
Publication Date
2013-10-02

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Abstract

The invention provides a RNA one-step-lysis and one-tube RT-PCR (one-Step-lysis and One-tube RT-PCR, SORT-PCR) detection method. Compared to the conventional RT-PCR method, the RNA one-step-lysis and one-tube RT-PCR method does not need to purify and extract RNA, RNA is hybridly combined to a fixed specific primer or Oligo primer after cell lysis, other components in the cell lysis solution supernatant are washed away, then the RNA is added into a RT reaction system to perform RT reactions. After being dried again, the RNA is added into a PCR reaction system to perform PCR reactions, and the result can be detected by the fluorescence real-time quantitative analyzing method. The RNA one-step-lysis and one-tube RT-PCR detection method shortens the RNA detection process of more than 3 hours to 2 hours, and has the advantages of simple operation, high detection sensitivity and specificity, and suitability for detecting RNA from various sources of sample lysis.
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Description

(1) Technical field

[0001] The invention relates to a one-step cleavage-tube RT-PCR detection method for RNA. (2) Background technology

[0002] RNA is the carrier of genetic information transmission. RT-PCR (reverse transcription–polymerase chain reaction) refers to a method that combines RT (Reverse Transcription, reverse transcription) and PCR (Polymerase Chain Reaction, polymerase chain reaction) reactions. RT-PCR uses RNA as a template cDNA synthesis, or RT, combined with PCR provides a fast and sensitive method for analyzing gene expression. RT-PCR detection or quantitative analysis of RNA is more sensitive and easier to operate than other RNA analysis techniques including Northern blotting, RNase protection analysis, in situ hybridization and S1 nuclease analysis. It is a powerful means for detection and analysis, detection and analysis of pathogens, etc.

[0003] The template for RT-PCR can be total RNA or poly(A)+RNA. RT requires RT enzymes (such as MMLV, AMV, e...

Claims

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