Posted 13 August 2012 - 02:33 AM
I have one question, i kept my pcr mix including my primer ,template with nuclease free water in the freezer at -20, and will amplify next day.. will it work???? or i have done a mistake icubating it, as taqmaster mix got finished and i have ordered it to make one more pcr.. kindly tell me i am in great tension here..
Posted 13 August 2012 - 03:36 AM
Posted 13 August 2012 - 05:19 AM
thanks a lot but I think i will be keeping it for 2-3 days at - 20. waiting for my polymerase to come.. i hope it works....
You anyhow you freeze all the components separetely for storage, don't you? So why woulnd't they work when they are together frozen? I did it before and it worked perfectly. Worst case scenario: the Taq polymerase in the mix will lose a bit of its activity.
Posted 13 August 2012 - 05:22 AM
Posted 13 August 2012 - 05:38 AM
thanks for ur reply but i froze it with polymerase, primer , abd my template, actually i had to do 4 pcr but eventually the taq master mix got over so I kept those 3 in freezer ...and waiting for the taq to come to make it for the another one.. do u still think i will be having proper amplification of my pcr.. m really nervous about that, and i think i have done a big mistake, already for the first time my pcr did not work...
It should work fine.
Posted 13 August 2012 - 07:41 AM
In the end of the day: it can work (most probably, if you did not do a big mistake the first time) or it will not work. You will know it only if you try. Otherwise, we can just talk whatifs until the end of time. I always say: while we talked here, you could have tried it already:)