I have been using folmer primer for the amplication of CO I gene in Caridean shrimps. But even though the DNA quantity is good, I'm not getting bands in AGE after doing PCR. I tried with dilution of DNA in 1/10 and 1/20 and put it on a gradient temperature PCR at an annealing temperature range of 47-55 degrees. Can anyone help me with this problem?
Try primer(COI) other than folmer. Sometime it happens to me also and I got good band when I used different COI primer.
@Vineesh Nedumpally thank you so much. But I have tried it with Crust folmer also. Are there any suggestions for the primer?
Sheena Jose "DNA barcoding Australia's fish species" please refer this article and try the primer listed here. May be it will help you.
Negative Results Decade of learning in marine DNA barcoding; the vertebrates,...
Dear Vineesh, May be before going for the PCR please check the primer in AGE first then try to go for Touchdown PCR. Less amount of DNA is good, Please see if it is a Universal primer having degenerate bases sometimes this happens. But please go for touchdown PCR it may help.
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