This kit contains U6 forward primer (U6 primer), and Universal 3'qPCR Primer that can be used for the quantitative detection of all miRNAs including U6. So only a specific forward primer for each target miRNA is needed for qPCR reactions. This Kit includes two steps: first, The RNAs are treated with gDNA Remover to eliminate the genomic DNA. Then, poly(A) tail is added to the 3’-end of miRNA molecules in the poly(A) tailing reaction. The reverse transcription reaction is performed using a specially designed Universal RT Primer to generate the first strand of the miRNA.

The Kit is a rapid reverse transcription kit containing high-efficiency DNase for the removal of genomic DNA. It contains fIve tubes of reagents: gDNA Remover, miRNA RT Enzyme Mix, 4× miRNA RT Buffer, Universal 3’qPCR Primer and U6 Primer.

Among them, gDNA Remover mainly includes concentrated DNase and buffer. It only needs to react at room temperature (19 ~ 27°C) for 5 minutes to degrade more than 95% of residual genomic DNA, which greatly reduces the interference to the results.

The miRNA RT Enzyme Mix mainly contains E. coli Poly(A) Polymerase, reverse transcriptase, and RNase Inhibitor. The E. coli Poly(A) Polymerase in it not only has high efficiency of adding poly(A) tail, but also specifically recognizes single-stranded miRNA, thus avoiding further reverse transcription reaction of miRNA precursor with double-stranded structure; The mutant M-MLV reverse transcriptase has strong anti-interference ability and amplification ability, and the amplification efficiency is particularly good.

The 4× miRNA RT Buffer contains all the raw materials and primers for miRNA polyadenylation and reverse transcription reaction, including Oligo (dT)-universal tag primers, buffer and dNTPs, and has been carefully optimized to ensure polyadenylation and the reverse transcription are simultaneously performed efficiently.