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primer regarding

Dear all,

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

Tags: confusion, primer

Replies to This Discussion

Permalink Reply by sambashiva daravath on April 1, 2011 at 4:09am

dear friend,

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

Permalink Reply by C.Vadivalagan on April 1, 2011 at 7:58am

Thank you friend but i need more about COI primer

Permalink Reply by Gaikwad Swapnil Sopan on August 5, 2011 at 4:05am

Dear Vadivalagan,

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.

Permalink Reply by C.Vadivalagan on August 6, 2011 at 3:35am

Dear Gaikwad,

thank you sir your valuable suggestion but i want specific primer sir if any idea please give me prime and PCR condition. that will be very useful for my research work

thank you

Permalink Reply by Gaikwad Swapnil Sopan on August 6, 2011 at 6:47am

Dear Vadivalagan,

Use the primers and conditiond mentioned in the paper. Following is the link for the paper. attached is the paper.

http://www.pnas.org/content/101/41/14812

Best of luck,

Keep posting.

Swapnil

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