my primer f27- 5' GAGTTTGATCCTGGCTCAG r1492-5'GGTTACCTTGTTACGACTT
My PCR doesn't answer but DNA extract was posivtie and i saw the bands
1. Are the orientation of these primers correct?
2. DNA template might have inhibitors, try to dilute them and do PCR again.
3. Do you have sample for a positive control? If so, include one.
Actinomycetes generally have high GC content genome. You can try adding DMSO to the reaction mix or you can use the Q5 DNA pol which always gave a positive PCR in our lab for actinomycetes.
Dear Dr.Yuan-Yeu Yau ·
1-ya its correct
2-we do diluted up to 1/100 then just one of them PCR was positive. and when i repeat the electrophoresis the only band which was observed once was not run second time.
3- we used the only sample which has band after PCR as positive control but when we repeat the electrophoresis it didn't make a band again
i confused what is happening again!
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