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storing large amounts of PCR mastermix beforehand


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#1 claritylight

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Posted 10 August 2009 - 07:02 AM

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?

#2 cellcounter

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Posted 10 August 2009 - 11:17 AM

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!

#3 jiajia1987

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Posted 10 August 2009 - 06:03 PM

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. =)

#4 ivanbio

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Posted 11 August 2009 - 07:54 AM

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.

Ivan
Carlsbad, CA




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