This product is a one-tube master mix for genomic elimination and reverse transcription, which can achieve genomic DNA removal and cDNA synthesis at the same time in a single-step operation, effectively avoiding the risk of sample contamination and RNA degradation caused by complex additions. The product contains a highly potent RScript II reverse transcriptase, which synthesizes the first-strand cDNA from very low amounts of total RNA or poly(A) mRNA, making it suitable for the reverse transcription of RNA templates with high-GC contents and complicated structures. The enzyme has greatly improved thermal stability, able to tolerate the reaction temperature up to 65°C and achieve a higher yield of synthesized cDNA. With the addition of the heat-sensitive dsDNase, the residual genomic DNA can be removed from the sample at room temperature.  The inactivation at 50°C does not affect the cDNA. The optimized 5×RScript II All-in-One RT Buffer is premixed with the Random Primer and Oligo18 (dT), compatible with genomic digestion and cDNA synthesis. Reverse transcripts are compatible with dye and probe qPCR for subsequent gene expression analysis.

• Performance

Figure 1. Reverse Transcription Performance of RK003-0050
Using 100 ng, 10 ng, 1 ng Hela cell RNA as templates, RK003-0050 was compared with two competitors in reverse transcription reactions followed by qPCR. Amplification curves showed excellent performance, especially at low template concentrations, surpassing competitors.

Figure 2. Genomic DNA Removal Performance of RK003-0050
Using 100 ng Hela cell RNA as the template, RK003-0050 was employed for reverse transcription, followed by cDNA synthesis and qPCR. The amplification curve indicates that RK003-0050 effectively eliminates genomic DNA, ensuring the accuracy and precision of the amplification reaction, and preventing interference from residual genomic DNA in quantification results.