PCR method capable of eliminating non specific braid and RT-PCR chip prepared using said method

Abstract

A PCR method for eliminating the non-specific band and precisely obtaining target gene by one step, and the RT-PCR chip prepared by said method are disclosed. When the primer of (PCR CRT-PCR or PCR) target gene is designed, the one or more bases at terminal 3' of primer are modified by thiophosphate. The polymerase used for extension reaction is the DNA polymerase with high Pfu fidelity. As a result, in various PCR reactions, only the template fully matched with 8bp at terminal 3' of primer can be extended with high fidelity to said template.

Description

technical field [0001] The present invention relates to a PCR (RT-PCR, PCR) method suitable for chip technology to obtain specific gene expression (including coding region, transcriptome, expression profile comparison), diagnosis and RT-PCR chip prepared by the method, especially A PCR method that can eliminate non-specific bands, accurately obtain the target gene in one step, and the RT-PCR chip prepared by this method to construct a high-precision, high-throughput method for obtaining specific gene expression (including coding regions, transcriptomes, etc.) , expression profile comparison), diagnostic technology platform. Background technique [0002] With the completion of the draft human genome DNA sequence ahead of schedule, human beings have entered the post-genome era. In the post-genome era, it is first required to use human genome DNA sequence information to study the structure and function of genes and their protein products, and further to ...

AI Technical Summary

Problems solved by technology

Therefore, the primer extension reaction mediated by Pfu high-fidelity DNA polymerase alone will also appear non-specific bands, which is also a defect that this type of high-fidelity DNA polymerase cannot be widely used

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