I am attempting to use 16s rRNA primers to amplify bacterial DNA for sequencing. Does anyone have experience using pcr to obtain amplicons? I am worried my thermocycler settings are not optimized...
Rene Christena
hi bonnie,
it depends on what pcr mix you are using, exactly follow the concentrations given in the instructions...specially primers and templates; to avoid unspecific binding or primer dimers...for my reactions i used promega go green mastermix..its works pretty well. 10uM of primers and 5uM of template would be enough.
all the best!!
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Grant Gallagher
Bonnie - relax - this is done all the time everywhere. Take a wander down the corridor to your local friendly micro lab and you'll get the answer.
Watch out for cross-contamination once you start though.
Good Luck
G
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