Validation of Custom Sequencing Primer

[Akira]

Hi.

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.

[giampe]

Hi,
maybe if you take a look into http://molbiol.ru/wiki/Cluster_generation_on_cBot
probably it will help to answer your question.