I've been having some problems with the PCR reaction... tested primers, water, PCR buffer, Taq and nothing came out so far. I'm guessing the next step should be testing the dNTP integrity, since we started having problems when the new stock arrived... any idea on how can I do it?
go back to somebody's supply of old NTPs and run parallel pcrs. I have never had a problem with NTPs but if they are made up much too strong then Mg will be removed so you could try upping the amount of Mg in the pcr or making up new NTP dilutions and running a pcr in parallel with only the new and old NTPs being different
go back to somebody's supply of old NTPs and run parallel pcrs. I have never had a problem with NTPs but if they are made up much too strong then Mg will be removed so you could try upping the amount of Mg in the pcr or making up new NTP dilutions and running a pcr in parallel with only the new and old NTPs being different
Thank you Paul Rutland, once again your advices were really usefull!!
Hi Claudia, one additional step you can also adopt that upon receiving any new lot of dNTPs just aliquot them in certain volumes you may need later, and use one by one as the dNTPs are quite sensitive to freeze thaw cycles, a major cause of their integrity loss.
Regards...
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