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Old 07-20-2012, 12:28 AM   #1
Location: Malaysia

Join Date: Sep 2010
Posts: 16
Default Validation of Custom Sequencing Primer


I would like to use my own sequencing primer. I design it with properties (Tm
and GC content and length etc) close to standard Illumina Sequencing Primer.

To check its performance I set up a PCR reaction with the primer as forward and P7 primer as reverse. But I don't know what is the annealing temperature I should use.

Anyone know what is the temperature for Illumina Sequencing Primer to anneal during cBOT last step (not sure if it's correct)? So that I can follow the temperature in my PCR step to see if my primer can anneal.

Please help. Thanks

- NGS newbie.
Akira is offline   Reply With Quote
Old 11-05-2012, 10:22 AM   #2
Location: Bari, Italy

Join Date: Aug 2009
Posts: 22

maybe if you take a look into
probably it will help to answer your question.
giampe is offline   Reply With Quote

annealing temperature, cbot, custom, illumina, sequencing primer

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