plz save my PCR product ! - (Feb/23/2008 )
my PCR was turned on and everything was ok with it..
unfortunately,,,the electrical current was interrupted
what shall i do ???
shall i put my PCR tubes on 4C ( freezer ),and then repeat the PCR ????
Just turn it on and run the PCR with the remaining number of cycles. The DNA is still there, so are other ingredients.
I don't know if however if it waits overnight, the polymerase might fail...
If the template matters run the final extention step to inactivate TAQ and store at 4oC. Use the products as templates for new PCRs. If it doesn't matter better start from scratch with new DNA.
thanks for your reply
but it came a little bit late
i took the tube and put it in the freezer
it took 1.30 hrs for the current to return
it is still in the freezer now...
what are your suggestions ???
am using it as molecular diagnosis for a mutation in a PATIENT
what is the difference ??
I think there is no harm to proceed... you have added all the ingredients after all.
run a gel later and you'll know. if there is a band, then congrats.
anyway, 1.30 hrs should be OK.
I used to have PCR running (overnight) in the machine and power supply was interrupted in the middle of the night. The PCR cycle resume after power supply came back. It was still OK, but the band looks slimmer.