Thermo Fisher Scientific
SuperScript™ II Reverse Transcriptase
18064014
Quantitative gene expression requires an efficient reverse transcriptase of mRNA. SuperScript™ II Reverse Transcriptase kit has an efficient recombined MMLV reverse transcriptase with minimal RNAse activity thereby increasing the processivity of the enzyme many folds up to 12kb. RNA stand is left along with the cDNA stand which can be degraded using the RNAse.
Cloning /quantitative gene expression
Total RNA
1 microgram of total RNA is taken as the starting material to which 1 microliter of oligo DT and 1 microliter of 10 micromolar dNTPs were added. Then makeup this to 12 microliters using nuclease-free water and heat to 65 degrees for 5 minutes followed by adding 4 microliters of 5x buffer and DTT were added and incubated for 2 minutes at 42 degrees. Finally, 1 microliter of MMLV is added and incubated at 42 degrees for one hour.
Good quality RNA should be used.
A good quality of cDNA is obtained at the end of the reaction which can be used for quantitative real-time expression or semi-quantitative PCR for cloning.
N/A
Easy to use
No drawbacks
If you are looking for an efficient cDNA conversion kit you can definitely choose this kit.