So I was doing a truseq DNA prep for a bunch of samples. I did PCR on the libraries and then did a gel extraction. The gel was a 2% agarose gel which I ran at 50 volts for about an hour and a half. I then did extractions of 180bp, 350bp and 600bp. I used a sigma gel extraction kit to purify the DNA and then ran the samples on a high sensitivity chip. I qubitted and got between 1.5 and 2ng/ul for all the samples. I got this very bizarre read for nearly all my samples. It's this fuzzy, ladder-like read. So I figured that there was some sort of contamination in my sample. I did a bead purification and ran a second HS bioanalyzer chip and got the strangest bands. In theory, it should have been clean bands, but I got anything but clean bands. I've included my initial gel extraction. It's the best gel extraction I've done. And then the pre-bead bioA run and the post-bead bioA run.
If anyone knows why I have these results, I'd really appreciate your help. Thanks!
If anyone knows why I have these results, I'd really appreciate your help. Thanks!
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