Hello all -
We've prepared our first library following a protocol originally for the Illumina Genome Analyzer. It's out for QC now and I'm keeping my fingers crossed. I'm now wondering, though, would it be possible to run our single-end library on the MiSeq? Is MiSeq only paired end or does it require a special P2?
The library is prepared using modified Solexa adapters. The second adapter (P2) is a divergence Y-type adapter so only fragments with P1 ligated first will amplify. Here are our adapter sequences:
P1:
P2:
which becomes, after PCR fill-in:
Will this single-end library work on a MiSeq? If you can explain why or why not, I'd be very grateful. Thank you in advance for any help.
We've prepared our first library following a protocol originally for the Illumina Genome Analyzer. It's out for QC now and I'm keeping my fingers crossed. I'm now wondering, though, would it be possible to run our single-end library on the MiSeq? Is MiSeq only paired end or does it require a special P2?
The library is prepared using modified Solexa adapters. The second adapter (P2) is a divergence Y-type adapter so only fragments with P1 ligated first will amplify. Here are our adapter sequences:
P1:
Code:
[FONT="Courier New"]Top: 5'- AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT-3' Bottom: 3'- AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT-5'[/FONT]
Code:
[FONT="Courier New"]Top: CTCAGGCATCACTCGATTCCTCCG[COLOR="Silver"]AGAACAA[/COLOR] Bottom: ACTCAGGCATCACTCGATTCCTCCGTCGTATGCCGTCTTCTGCTTG[/FONT]
Code:
[FONT="Courier New"]Top: CTCAGGCATCACTCGATTCCTCCGTCGTATGCCGTCTTCTGCTTG Bottom: ACTCAGGCATCACTCGATTCCTCCGTCGTATGCCGTCTTCTGCTTG[/FONT]
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