Appreciate any help on this.
I am following the Genomic Dna Library protocol from Solexa. In the final step (step 6) which amplifies the adapter enriched products from the gel by pcr, Solexa uses Phusion Dna Polymerase. For some reason, this step does not always work for me as I do not get any pcr bands in my region (250bp). I did the first few times with my control, but it does not always work with my actual samples. I will increase my cycles from 18 to 22 and see what that does, but are there alternatives I can try to the Phusion DNA polymerase? Will any DNA polymerase suffice? I was thinking of using the Taq DNA polymerase I use in my other PCR (non-solexa related) and I know works. Have others have tried using polymerases besides the Phusion that solexa provides?
I am following the Genomic Dna Library protocol from Solexa. In the final step (step 6) which amplifies the adapter enriched products from the gel by pcr, Solexa uses Phusion Dna Polymerase. For some reason, this step does not always work for me as I do not get any pcr bands in my region (250bp). I did the first few times with my control, but it does not always work with my actual samples. I will increase my cycles from 18 to 22 and see what that does, but are there alternatives I can try to the Phusion DNA polymerase? Will any DNA polymerase suffice? I was thinking of using the Taq DNA polymerase I use in my other PCR (non-solexa related) and I know works. Have others have tried using polymerases besides the Phusion that solexa provides?
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