I am preparing a DNA library from soil microbial DNA samples using the KAPA HyperPlus kit. After a final size selection using their protocol (.7X cut followed by .9X cut with KAPA Pure beads), I ran a bioanalyzer chip and seem to have two separate DNA fragments peaks, one with quite large DNA fragment sizes. Subsequent clean-ups don't seem to get rid of the second peak, I just lose DNA across the board. Any ideas?
Hi Marla, looking at the 11 sample pictures you provided I imagine your issue is bead carryover. Be careful with your pipetting during the elution step and avoid disturbing beads. You can also try a different magnet if you have one to see if that makes a difference. Do you have a negative control (one with no DNA sample) to see if this artefact contamination also occurs here?
Thank you for the suggestion! I will try a negative control on my next run. I have been having trouble with part of the bead pellet sliding down the tube when drawing off eluate, but thought I was being careful not to get any in the pipette. Maybe not careful enough...
- Badges
- Science topic
- Similar topics
- Biological Science
- Histology
- Sectioning