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03-01-2011, 08:50 PM
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#1 |
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Junior Member
Location: Korea Join Date: Oct 2010
Posts: 4
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Non-specific band after gel purification of PCR products
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. |
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03-02-2011, 09:24 PM
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#2 |
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Junior Member
Location: Ames, IA Join Date: Nov 2010
Posts: 2
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You might try a nested PCR reaction on the gel-purified product
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10-21-2013, 10:44 PM
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#3 |
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Junior Member
Location: Japan Join Date: Sep 2012
Posts: 2
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any progress omnivore? I got the same problem now. ty. the sequencing worked fine tho.
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04-05-2017, 07:19 PM
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#4 |
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Member
Location: USA Join Date: Jul 2015
Posts: 28
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It will not affect your seq, or limited affect. It may be a dimer of your product.
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| Tags |
| amplicon, gel purification, non-specific band, pyrosequencing |
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