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

The present invention discloses a method to eliminate non-specific bands, accurate one-step PCR method to obtain the desired gene. In particular: the gene of primer PCR (RT-PCR, PCR) design, primer 3 'end of one or more bases using phosphorothioate modified; the polymerase extension reaction with Pfu DNA polymerase fidelity enzyme. Because most 3'-5 'exonuclease activity of the Pfu DNA polymerase fidelity can be identified and close the primer 3' end of the extension reaction 8bp any one or more mismatched bases. Therefore, various PCR reactions, only 'end of the template can be extended 8bp exact match primers 3 and fidelity to maintain a height extending in the template. The present inventors constructed a versatile, high-precision, high throughput, chip technology applicable to obtain specific gene expression (including coding regions, transcriptome, comparing expression profiles) in diagnostics and technology platform.