Hello,
If I use a PCR plate for library prep and I need the products after each step in the protocol (in my case it's polyadenylation, RT and PCR), is it safe to open the plastic foil that covers the plate without disturbing the liquid so that some drops 'jump' between wells? Also, the I guess it is hard to keep the wells into which the matermix is not dispensed covered at any given time.
Perhaps the best way to optimize workload and contamination is to work with PCR strips, have the mastermix aliquoted to 96 wells, and then with a multi-channel pipette dispense master mix for to one strip (containing 8 tubes), while all the other strips are closed. You open the strip you want to dispense the mix into just for a few seconds, thus reducing the chance for contamination. Does it make sense?
Sorry if my questions seem weird...
Iddo
Hello Iddo,
I've worked with both plates and microplates for qPCR reactions and yes, for preventing contamination it would be good if you use strips and go as you say, working with 8 at a time. We had problems with some data we were trying to collect amplifying from gDNA a viral DNA strand, the best solution for avoiding contamination was to work with the strips, good luck with your work!
Hi
Yes, I also agree with Jose. Better use PCR strip or single tube if you want to handle library prep manually. My colleague also suggested to use dome cap instead of flat cap tube. Better handle small (but not too small) number of sample based on your capability and don't over do it rather trying to handle so many sample at a time. For me, the best I can handle is 6 at a time. I did get a good quality data. All the best to you!
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