I am conducting my Final Year Project which involves PCR, however, due to time constraints caused by clashes in my schedule, I can only run PCR the next day. In order to optimise time, can I mix Go Taq Green Mix, Nuclease Free Water & DNA template on Day 1 & store them in the freezer overnight? And subsequently, on Day 2, add my forward & reverse primers to the mixture and place my samples in the PCR machine.
If possible, are there any articles you can share about this?
You can try. Close cap tightly to prevent evaporation. And if you have multiple samples then open very carefully to avoid contamination. Usually template and Taq are added at last.
You can make up mastermixes in advance of running ther assay because taq is very thermostable. I would not store the mix in a freezer because enzymes do not like being frozen and thawed ( taq comes in glycerol containing storage buffer so that it does not freeze) but it will keep at 4c for days and retain its activity
Mixing the ingredients on the same working day is better to avoid the issue of the preferred temperature difference for each ingredient
You could, but in my experience, mixing the reagents is the fastest step. It's honestly not worth it. You'll spend more time writing down your experimental design, labeling and pipetting your template than you would mixing the enzyme + primers + etc.
If you are testing the same DNA with a bunch of different primers, it might make sense to make a plate with aliquots of all of the template DNA + controls that you are testing and use a multichannel to add DNA to your reactions. That is a reasonable way to save time.
Note: do NOT add DNA to your enzyme mix! You have to include a proper set of controls, including a reaction where you add water instead of DNA. Otherwise you won't know if you have contamination. Trust us on this one.
Good luck!
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