StarLighter reverse transcription super premix (including gDNA removal) is developed based on the new generation of reverse transcriptase (the fourth generation of genetic engineering enzyme). The Mix contains the reverse transcriptase, random primers, Oligo dT and other reaction components required for reverse transcription. The RNA template can be directly added to the system for reverse transcription reaction. It is also equipped with gDNA Removal and Buffer for efficient removal of gDNA, which can effectively remove gDNA contamination during reverse transcription. The new generation of reverse transcriptase is resistant to high temperatures and can show good reverse transcription activity at 42°C-65°C, and can effectively reverse transcribe RNA with complex structures.
1. High reverse transcription efficiency, accurately reflecting template information
2. High temperature resistance and effective reverse transcription of complex RNA
3. Remove gDNA contamination in one step without affecting reverse transcription performance
4. Compatible with high GC/AT templates and degraded samples, easy to operate
1. Efficient removal of gDNA
1.1 Mouse Total RNA with different starting amounts (0.1ng, 1ng, 10ng, 1000ng) containing 10% gDNA contamination was treated with gDNA and reverse transcribed, followed by qPCR quantification to evaluate the gDNA removal ability of StarLigher Reverse Transcription Super Mix (with gDNA Removal) and the reverse transcription performance of RNA under gDNA contamination. The results are as follows:
1.2 gDNA removal and reverse transcription were performed on 100ng mouse Total RNA containing different amounts of gDNA contamination, followed by qPCR quantification to evaluate the gDNA removal ability of StarLigher Reverse Transcription Super Mix (containing gDNA removal) and the reverse transcription performance of RNA under gDNA contamination. The results are as follows:
2. Reverse transcription quantitative results accurately reflect the template amount information
RNA input and qPCR quantitative output showed an excellent linear relationship. Different reverse transcription reagents were used to reverse transcribe mouse Total RNA with different starting amounts (0.1ng, 1ng, 10ng, 1000ng), followed by qPCR quantification. The linear relationship between RNA input and qPCR Ct value of different reverse transcription reagents was compared. The results are shown in the following figure:
