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Old 06-06-2011, 09:35 AM   #1
ZAAB
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Location: Boston

Join Date: Apr 2011
Posts: 18
Default Library PCR enrichment questions (with image!)

Hi All:
I'm preparing libraries (five of them)...all are PCR products to begin with...so size is very uniform, and I avoided the fragmentation and end repair. I picked up at the A-tailing, moved forward to ligating adapters, gel purified 'em, and moved on to PCR.

Attached is a gel showing sets of five:

First set: Starting material (insert).
Second set: After ligation of adapters, gel purified (thought I did a good job).
Third set: after 10 rounds of PCR.

Ladder is 766, 500, 350, 300, 250, 200(bright), 150, 100, 75, 50, 25.

My Questions: What are the large bands? I can see my correct bands within the haze, floating at ~300. Should I gel purify again? Anyone seen this before?

Thanks.
ZAAB
Attached Files
File Type: pdf ZN06062011_01.pdf (466.9 KB, 356 views)
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