Posted 13 January 2011 - 09:31 AM
Primers do not go bad all in one go. In fact, I have not actually seen a case where the fault is the primers.
And from what you have written, you do have stock primer solution and a diluted working primer solution.
Usually when a PCR suddenly doesn't work it is the fault of the template. Then the dNTP, then the thermocycler....
I would suggest that you run the PCR on a sample of DNA that you know will give you a signal.
I have seen peltier of a thermocycler stop working once. You might want to note where on the heating block did you place your tubes. Find out if anybody complained about PCRs not working
I saw it twice... 2 of the Biorad Mycycler in my collaborative lab had stop working more than half year ago... the repair almost cost about purchasing a new machine.
Be sure to check the running report everytime after run.
Expecting the world to treat you fairly because you are a good person is like expecting the lion not to attack you because you are a vegetarian.
..."best of our knowledge, as far as we know this had never been reported before, though I can't possible read all the published journals on earth, but by perform thorough search in google, the keywords did not match any documents"...
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