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-   -   help with TruSeq (http://seqanswers.com/forums/showthread.php?t=11907)

upendra_35 06-08-2011 10:14 AM

help with TruSeq
 
1 Attachment(s)
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

pmiguel 06-08-2011 10:27 AM

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

mnkyboy 06-08-2011 11:03 AM

You might want to try repurifying them with the beads.

upendra_35 06-08-2011 11:27 AM

1 Attachment(s)
Hi pmiguel, marker sizes added now.


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