DIY illumina adapters

ZAAB · 05-03-2011, 07:00 AM

Hi All:
Trying to determine whether I can get away with making my own adapters for the HiSEQ 2000.
I've got a library of small DNA (125nt) fragments, and would like to sample the diversity of this library pre- and post-selections.

I get it that there is that funky fork with a Phosphorothioate Bond...and that the Tm of the sequencing primer is in the high 70s. Has anyone built their own adapters, or should I just buy the genomic DNA prep kit...?

Also, I am not compiling huge contigs, but rather, assessing the diversity of the library through selections...what, other than the QC descriptions, do I need to assess this diversity? Can someone point me towards the PERL scripts that would process these data into counted bins of reads?

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05-03-2011, 07:00 AM	#1
ZAAB Member Location: Boston Join Date: Apr 2011 Posts: 17	DIY illumina adapters Hi All: Trying to determine whether I can get away with making my own adapters for the HiSEQ 2000. I've got a library of small DNA (125nt) fragments, and would like to sample the diversity of this library pre- and post-selections. I get it that there is that funky fork with a Phosphorothioate Bond...and that the Tm of the sequencing primer is in the high 70s. Has anyone built their own adapters, or should I just buy the genomic DNA prep kit...? Also, I am not compiling huge contigs, but rather, assessing the diversity of the library through selections...what, other than the QC descriptions, do I need to assess this diversity? Can someone point me towards the PERL scripts that would process these data into counted bins of reads?