Hi all, I've run into a minor problem doing Illumina sample prep + exome enrichment. Everything worked, except when I ran the samples on a Bioanalyzer at the end, I got a peak way out at like 3kb, in addition to the peak I wanted (300-400bp). There's also a tiny hump at about ~650bp, but it's small enough that I'm just going to ignore it.
I'm using Kapa Library Amplification Kits for the first time, so I presume I'm just not optimizing the conditions correctly. I don't think it's contamination, as it's occurring in all of my samples and it's getting amplified in PCR, so it must have adapter sequences in it somewhere, and no-one else in the lab was handling sequencing libraries at the time. Can anyone suggest what might have caused this, and what the high weight DNA could be? Also, what should I do about it? Ignore it, size-select it out, or ditch all my samples and start over?
I'm using Kapa Library Amplification Kits for the first time, so I presume I'm just not optimizing the conditions correctly. I don't think it's contamination, as it's occurring in all of my samples and it's getting amplified in PCR, so it must have adapter sequences in it somewhere, and no-one else in the lab was handling sequencing libraries at the time. Can anyone suggest what might have caused this, and what the high weight DNA could be? Also, what should I do about it? Ignore it, size-select it out, or ditch all my samples and start over?
Comment