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as i am doing my doctoral research in DNA barcoding on selected butterflies in south western ghats, India.I completed my field work of 1st chapter. now i started my isolation and PCR regarding the experiment i have some doubt,
1. I have to sue of DNA isolation Kit (because of proper result)
2. PCR Process
COI F - GGA GGA TTT GGA AAT TGA TTA GTT CC
COI R - TCC AAT GCA CTA ATC TGC CAT ATT A
but here how to identified the total bp of following primer and PCR setup like temperature cycles and etc.
i tried lot of time but i didn't get proper way kindly help me fallowing questions
the size of the COX 1 is nearly 600-700 bp. the above designed primers are may attach with in the coI region it self only. the PCR conditions will be normal as we do in laboratory. ok
all the best bye
Form which paper you are referring this primer ? Usually in the paper conditions are mentioned and if it is lep F1 and Lep R1, it gives amplification, Try out this 95- 5min, 35 cycles of 94- 30 sec, 51- 30 sec, 72- 1 min and final extension at 72- 5 min.
Use the primers and conditiond mentioned in the paper. Following is the link for the paper. attached is the paper.
Best of luck,