Application of Bst DNA polymerase in RNA amplification

Abstract

The invention discloses an application of Bst DNA polymerase in RNA amplification. The Bst DNA polymerase has reverse transcriptase activity, and thus RNA is subjected to inverse transcription into cDNA under action of the Bst DNA polymerase. A new RNA amplification detection method is established on the basis of the new discovery that the Bst DNA polymerase has reverse transcriptase activity. The method integrates inverse transcription and subsequent nucleic acid amplification, temperature adjustment is not needed during the whole process, addition of extra reverse transcriptases is not needed, operation is simple, reaction time is shortened, and new methodology reference and technical support are provided for RNA inverse transcription and amplification detection. The discovered Bst DNA polymerase can perform an inverse transcription reaction at a high temperature, which facilitates to open and combine RNAs with complex secondary structures, and therefore the problem is solved that reverse transcriptases of AMV or the like are difficult to amplify RNA advanced structure areas.

Description

Background technique

[0001] RNA is the core and key to the expression of genetic information in the genome, and is also an important object of life science research. Although the purpose and technical means of RNA research are different, it is a common requirement to obtain sufficient amounts for follow-up research. With the application of many advanced experimental equipment and technical methods, there are more and more RNA amplification methods, but biomedicine, inspection and quarantine, etc. put forward higher requirements for rapid RNA amplification, making it more suitable for on-site testing Needs.

[0002] RNA amplification uses RNA as a template, reverse transcription into cDNA, and then amplifies on the basis of cDNA. At present, the commonly used methods for amplifying RNA include reverse transcription PCR (RT-PCR), rolling circle amplification (RCA), nucleic acid sequence-dependent amplification technology (NASBA), exponential strand replacement technology (exponenti...

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