3 replies to this topic

[claritylight]

Would it be fine to make up large amounts of PCR mastermix, but exclude Taq polymerase, DNA template, and primers? I want to store overnight in the freezer so I can just come in and start the next day or perhaps within that week. Just wondering the length of stability of the mastermix would be over time too?

[cellcounter]

claritylight, on Aug 10 2009, 08:02 AM, said:

Would it be fine to make up large amounts of PCR mastermix, but exclude Taq polymerase, DNA template, and primers? I want to store overnight in the freezer so I can just come in and start the next day or perhaps within that week. Just wondering the length of stability of the mastermix would be over time too?

I have stored such mastermixes (even including Taq) overnight at 4'C,  without any appreciable reduction in PCR products. I have not been so daring as to store it in -20 or for prolonged periods of time.

I guess it also depends upon the robustness of your product specific PCR. But let us know your insights if you try this out at your expense!

[jiajia1987]

I do not and have never stored any mastermixes for prolonged periods of time. I always prepare my mastermix just before I am going to do my PCR. I guess it shouldn't do any harm to store mastermix overnight at -20 though, so long as you ensure you spin down everything and mix them well before you use it for your PCR.

Do let us know how things go for you. =)

[ivanbio]

I think I may be missing something: if your PCR master mix does not contain Taq or primers, then why prepare it ahead of time? I guess you can mix the dNTPs and reaction buffer ahead of time, yet I do not see the advantage of this since you still would need to add the Taq and primers before using it.