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Old 06-08-2011, 10:14 AM   #1
upendra_35
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Location: USA

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Default 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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File Type: pdf TruSeq libraries 1-8.pdf (592.8 KB, 49 views)
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Old 06-08-2011, 10:27 AM   #2
pmiguel
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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.

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Phillip
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Old 06-08-2011, 11:03 AM   #3
mnkyboy
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You might want to try repurifying them with the beads.
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Old 06-08-2011, 11:27 AM   #4
upendra_35
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Hi pmiguel, marker sizes added now.
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