I was preparing the 2nd hybridization step of an exome sequencing project. But I mixed up the target elution buffer and the target capture buffer, and used the wrong one (i.e. I used the elution buffer to do the capture). In trying to get back the captured DNA sample, I extracted it with AMPure Beads and run a DNA 1000 chip with the sample. Unfortunately, from the result, I could not find any DNA > 200bp. There is only one sharp band with 20ng/ul at 100-150bp. I suppose that is the target capture oligo I put in the reaction for 2nd hybridization. I also saved the 'wash waste' from AMPure Beads, and found no DNA in it with DNA 1000 chips. I do not understand why AMPure Beads can save the oligos but not the captured library. Do you know if I have any chance I can find back my capture library?
On the other hand, I continue the 2nd hyb with the DNA I extracted from AMPure Beads, are they safe to be kept at -20C before the wash?
Thanks!
On the other hand, I continue the 2nd hyb with the DNA I extracted from AMPure Beads, are they safe to be kept at -20C before the wash?
Thanks!
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