This kit is a special kit for the first-strand cDNA synthesis by poly(A) microRNA (miRNA), which contains all the raw materials for the miRNA -poly A-tail and reverse transcription reactions. Through careful optimization, it ensures that the poly(A) modification process at the 3' end of miRNA and reverse transcription process can be carried out efficiently at the same time. The RScript II miRNA-A enzyme mix contains the poly(A) polymerase (PAP) and RScript II Reverse Transcriptase. The high efficiency rendered by the PAP-poly A tail can specifically recognize the single-stranded RNA and effectively avoid the reverse transcription reaction of miRNA precursors with the double-stranded or stem-loop structures. RScript II Reverse Transcriptase has been engineered to lose the RNase H activity and increase the affinity with RNA, resulting in a significant increase in the reverse transcription efficiency and sensitivity of miRNAs. The reverse transcription products obtained with this product can be directly used for subsequent quantitative detection by dye or probe methods.

Figure 1. Sensitivity Test of RK005-0050
RK005-0050 was tested with different concentrations (1000 ng, 100 ng, 10 ng, 1 ng, 0.1 ng) of mouse RNA as templates, targeting the Mmu-mir21-5p gene. Amplification curves demonstrated good sensitivity, reaching as low as 10 pg total RNA.

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Figure 2. Amplification Performance of RK005-0050 on Templates from Different Sources
(A) The hsa-U6 gene of Hela cell RNA
(B) The hsa-mir16-5p gene of Hela cell RNA
(C) The Mmu-U6 gene of mouse RNA
(D) The Mmu-mir24-3p gene of mouse RNA

RK005-0050 was used with templates from different sources and concentrations, showing excellent amplification performance for various target genes.