The EZBioscience® 4× Reverse Transcription Master Mix (with gDNA Remover) is a kit suitable for real-time RT-PCR (RT-qPCR) that contains a gDNA Remover which can effectively eliminate the contamination of genomic DNA (gDNA) or other double stranded DNA during the analysis of gene expression. In order to accurately analyze gene expression, it is necessary to detect cDNA in samples without contaminating DNA.
To avoid amplification of gDNA, primers can be designed on different exons spanning introns. However, there may be cases where a suitable primer cannot be designed, as with a gene with a single exon or a gene without a long intron. Also, it may be difficult to avoid unexpected amplification from gDNA due to non-specific amplification or the existence of pseudo-genes. Moreover, some labs are heavily contaminated by the PCR products of previous tests.
The kit offers a potent gDNA Remover that can eliminate double stranded DNA in RNA sample in 5 minutes at 25 °C without loss of RNA. Then first strand cDNA is synthesized by adding the 4× RT Master Mix and primers, and react at 42°C for 15 min.
Reaction products are applicable to subsequent PCR and qPCR. The mix contains Reverse Transcriptase, RNase Inhibitor, optimized buffer system, dNTP, and Oligo dT and Random Hexamer as primers. And gDNA Remover is supplied in a separate tube. All of the components should be stored at -20°C.
For detailed information, please check the product manual.