Greetings,
I am performing a reverse transcription using the taqman microRNA assay protocol.
Parameters of thermocycler for this step are as follows: 30 min/16°C, then 30 min/42°C, then 5 min/85°C.
The program was only running for about 8 minutes, then the power failed for about 5 minutes before returning, and the program was restarted again.
How would that affect my reverse transcription product?
What would be the best course of action if such event occurred again?
Thank you.
Greetings,
I wouldn't worry about it, it's only 5 minutes at 16 °C, the only cons I can think about is that, in order to standarize your protocol, you would have to change the conditions of your next samples to match your dephased samples. However, as long as you don't open the thermocycler lid, the temperature shouldn't change a lot, so no big deal there. If you want to follow the protocol to the Best you can, I'd recommend you to keep a water bath at 16 °C as a backup in case of this kind of accidents happen again, where you could put your samples for as Long time as you need, or even better, connect the thermocycler to a no-break.
Good luck
Nada Moustafa i believe nevertheless the power failure would have affected the cDNA synthesis process to a certain extent. Thus, I would suggest you carry out the cDNA synthesis again whenever possible. Also, try to avoid thermocycler machines that tend to experience power failure if possible.
Hello Nada
It happened to me a lot! I always do continue the program from where it stopped if the power failure was short as 5-10 min. My PCR machine is also a good one, it starts automatically from where the power failed.
Regards
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