My CDNA reaction was accidentally heated above 50C and the thermocycler went all the way to 70C before I immediately took it off the block and put it on ice. It was at 70C for about 2-3 seconds. Will the reverse transcriptase still work?
Depending on the RT you are using, at standard temp MLKV RT would last 5-10 at ~55 °C. At such a high temperature, I wouldn' t count on it. But, as long as you are working on a thermocycler, the temperature ramps very fast, you might hace a change. Some genetically engineered MLKV RTs would last for ~30 min, wich is the case for the majority of RT kits. However, I'd run the reaction from the begining, since the might be slight differences in the efficiency among your distinct Rx batches. Good luck.
Not wild MLKV RT Nadia, here' s a nice and interesting paper https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2632894/
Thank you Adán Valenzuela Castillo for sharing the paper....but still the half life is approx 5 min which is more than the 2-3 sec Chloe Thangavelu mentioned.
I was going to tell her that run a fresh reaction with this one so that she would know for sure, and can use it as a reference in future
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