Description
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 room temperature without loss of RNA. Then first strand cDNA is synthesized by adding the 4× RT Master Mix and primers. Reaction products are applicable to subsequent PCR, qPCR and PCR cloning. The template could be mRNA, microRNA, lncRNA, circRNA, etc.
The Reverse Transcriptase in this Mix is a genetic engineered enzyme based on M-MLV (RNase H-) reverse transcriptase. The reverse transcriptase lacking RNase H activity is suitable for preparing full-length cDNA. And the multiple site-mutations of Reverse Transcriptase can obviously increase its affinity to RNA templates and its strand extending ability, which make reverse transcription reaction more efficient. Moreover, this transcriptase is rather resistant to common reverse transcriptase inhibitors. This product is also very suitable for reverse transcription using plant tissue RNA.
Components
Components | A0010G (100 Rxns) | A0010G-L (500 Rxns) |
4× RT Master Mix | 550 μl | 550 μl × 5 tubes |
Oligo dT18 | 110 μl | 110 μl × 5 tubes |
Random Hexamer | 110 μl | 110 μl × 5 tubes |
gDNA Remover | 220 μl | 220 μl × 5 tubes |
Nuclease free ddH2O | 1 ml | 1 ml × 5 tubes |
Storage
Store at -20°C.
Features
1. Oligo dT18 and Random Hexamer in this kit are packaged separately, so different types of reverse transcription primers can be used flexibly for different experimental designs.
2. gDNA Remover can quickly and completely remove genome contamination, eliminating the influence of genomic DNA on the accuracy of qPCR results.
3. Extensive template compatibility: Compatible with RNA templates of different species and poor integrity.
For detailed information, please check the product manual.
