help with TruSeq

[upendra_35]

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]

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]

You might want to try repurifying them with the beads.

[upendra_35]

Hi pmiguel, marker sizes added now.