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Old 03-17-2019, 03:55 PM   #1
JVGen
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Join Date: Jul 2016
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Default Nextera Read 1/2 Primer Sequences?

Can anyone confirm the Read 1 & Read 2 primer sequences for libraries prepped by tagmentation with Nextera?

I recently asked Illumina if the following adapter sequence would work for a modified approach, and they said that it was truncated and missing part of the read 1 primer sequence (which reads from i5 towards the insert). From what I can tell, I've included everything except the Adapter sequence that is common to both i5 and i7 adapters. I didn't expect that to be part of the Read Primer, but I guess this could work if the primers bound to opposite strands?

Proposed Adapter Sequence:
5’-AATGATACGGCGACCACCGAGATCTACAC[i5-index]TCGTCGGCAGCGTC

Green is the P5 sequence that binds to the flow cell.
Blue is the sequence that binds to the adapter incorporated by Tn5 during tagmentation.

From Simone Picelli's 2014 paper, the Tn5 adapter sequences are:

Tn5ME-A
5'-TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG-3'
Tn5MErev5′
3’-OH-TCTACACATATTCTCTGTC-PO4-5’
Tn5ME-B
5′-GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG-3'
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