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Old 10-31-2013, 09:05 AM   #1
Location: UK

Join Date: Nov 2010
Posts: 49
Default lack of blocking primer in pre and post hyb...

It looks like I made a mistake in the hybridization library prep. Following the Rohland2012 protocol I forgot to use truncaded primers for pre-hyb (6 cycles). I used the fully indexed and flow cell ready primers for this step (no blocking primers added either)...
After the post-hyb amplification I was able to see a product, but after the sequencing was completed almost exclusively illumina adapters were present. Does it look like a showcase for daisy-chaining PCR product ?
What are my options with regards to library ? Can I simply go to post hyb library left over and use blocking primers ? Shall I go back before pre-hyb ?
Thanks for any help!
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Old 11-06-2013, 03:09 AM   #2
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Location: Amsterdam

Join Date: Nov 2009
Posts: 112

I would do the hyb again with the blockers.
Zaag is offline   Reply With Quote

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