Hi,
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!
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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