Description
The EZBioscience® Color Reverse Transcription Kit (with gDNA Remover) is a new-generation reverse transcription kit containing high-efficiency DNase for the removal of genomic DNA and high-stability blue dyes, which has higher reverse transcription efficiency than the previous generation.
To accurately analyze gene expression, it is necessary to detect cDNA in samples without contaminating DNA. To avoid amplification of genomic DNA (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 is suitable for real-time RT-PCR (RT-qPCR) that contains a gDNA Remover which can effectively eliminate the contamination of gDNA or other double stranded DNA in RNA samples at room temperature for 5 minutes without loss of RNA during the analysis of gene expression. Then the first strand of cDNA is synthesized by adding the 4× RT Master Mix and primers. Reaction products are applicable to subsequent PCR, qPCR and PCR cloning. The RNA sample could be mRNA, microRNA, lncRNA, circRNA, etc.
The 4× RT Master Mix of the kit contains reverse transcriptase, RNase Inhibitor, optimized buffer system, dNTPs, and blue dye. blue dye in the 4× RT Master Mix could avoid of pipetting errors. It can be used with the EZBioscience® Color SYBR Green qPCR Mix (A0012, A0012-R1, A0012-R2), which is supplemented with an inert red dye. Mixing cDNAs, primers and other components with the EZBioscience® Color SYBR Green qPCR Mix together in a qPCR reaction turns the solution into purple, which provides a visual aid when pipetting and decreases the risk of pipetting errors during reaction setup, especially when using white reaction vessels. And the dyes do not affect the specificity or sensitivity of qPCR reactions. So, these two kinds of kits are recommended to use together to get optimal results.
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. At the same time, this kit uses the latest optimized reaction system to further improve the reverse transcription efficiency. This product is also very suitable for reverse transcription using plant RNA.
Components
Components | A0010CG (100 Rxns) | A0010CG-L (500 Rxns) |
gDNA Remover | 220 μl | 220 μl × 5 tubes |
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 |
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. 4× RT Master Mix contains blue dye, which can be used with the EZBioscience® Color SYBR Green qPCR Mix (A0012, A0012-R1, A0012-R2), which is supplemented with an inert red dye. Mixing cDNAs, primers and other components with the EZBioscience® Color SYBR Green qPCR Mix together in a qPCR reaction turns the solution into purple, which provides a visual aid when pipetting and decreases the risk of pipetting errors during reaction setup, especially when using white reaction vessels.
4. Excellent reverse transcription efficiency: Compared with common commercial reverse transcription products, this kit reverses cDNA with the smallest quantitative CT value and has excellent reverse transcription efficiency.
5. Extensive template compatibility: Compatible with RNA templates of different species and poor integrity.
For detailed information, please check the product manual.
