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06-08-2011, 11:14 AM
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#1 |
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Senior Member
Location: USA Join Date: Apr 2010
Posts: 102
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help with TruSeq
I have recently used TruSeq kit for making 8 of Brassica RNAseq libraries and after amplification (15 cycles) and running on the gel (1% Agarose) i found that 5 out of 8 libraries have a broader range of fragments (smear) compared to the rest of the 3 libraries. I don't know why and the only reason i can think of possible over-amplification. But if it is so why only 5 and not all 8 have this problem. Any help is appreciated.
I have attached the picture of the gel for easy understanding. Thanks |
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06-08-2011, 11:27 AM
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#2 |
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Senior Member
Location: Purdue University, West Lafayette, Indiana Join Date: Aug 2008
Posts: 2,317
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The picture you attach looks to my viewer (FoxIt pdf viewer) like it has been processed through some bizarre filter to make it look like a scanning electron micrograph. That is off-putting -- but also you don't give the marker sizes.
-- Phillip |
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06-08-2011, 12:03 PM
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#3 |
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Member
Location: Seattle, WA Join Date: Mar 2009
Posts: 87
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You might want to try repurifying them with the beads.
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06-08-2011, 12:27 PM
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#4 |
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Senior Member
Location: USA Join Date: Apr 2010
Posts: 102
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Hi pmiguel, marker sizes added now.
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