Hello!
My research right now involves sequencing DNA sequences that were amplified by degenerate primers in PCR. I have been informed by Laragen that it's not possible to sequence my PCR amplicons because of the degeneracy at the beginning and end of each sequence. So I have had to use TOPO clone libraries to get M13 sequences incorporated at the very beginning and ends of my amplicons. Using this approach, I have gotten sequence results, but I am wondering if I am overlooking a possibly easier protocol.
Thanks in advance for your replies.
My research right now involves sequencing DNA sequences that were amplified by degenerate primers in PCR. I have been informed by Laragen that it's not possible to sequence my PCR amplicons because of the degeneracy at the beginning and end of each sequence. So I have had to use TOPO clone libraries to get M13 sequences incorporated at the very beginning and ends of my amplicons. Using this approach, I have gotten sequence results, but I am wondering if I am overlooking a possibly easier protocol.
Thanks in advance for your replies.
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