I'm trying to purify my PCR products (obtained by using barcoded fusion primes) by gel purification to increase the quality of amplicon pyrosequencing.
My problem is that non-specific band of about 1200 bp is not removed by gel purification process (please see the attached pdf file. My product size is about 600 bp and I usually cut out gel slices between 550 and 650 bp).
Do you know how it can occur? And, If I run these samples without further purification, how badly does the non-specific band affect the overall sequencing results?
Thanks in advance.
My problem is that non-specific band of about 1200 bp is not removed by gel purification process (please see the attached pdf file. My product size is about 600 bp and I usually cut out gel slices between 550 and 650 bp).
Do you know how it can occur? And, If I run these samples without further purification, how badly does the non-specific band affect the overall sequencing results?
Thanks in advance.
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