Hi guys-
Using the new illumina small RNA sample prep kit v1.5, do we expect an additional "A" right after the miRNA sequence?
Here is the original 3' RNA Adaptor sequence and result library and I added the red colored "A":
3' RNA Adapter
5' P-AUCGUAUGCCGUCUUCUGCUUGUidT
Result Library:
5' AATGATACGGCGACCACCGA CAGGTTCAGAGTTCTACAGT CCGACGATC (N) ATCGTATGCCGTCTTCTGCTT G 3'
3' TTACTATGCCGCTGGTGGCT GTCCAAGTCTCAAGATGTCA GGCTGCTAG (N) TAGCATACGGCAGAAGACGAA C 5'
I suspect that illumina improved the new kit using pre–5',5'-adenylated adaptor, so the adaptor ligation can be performed in absence of ATP. However, I am not able to find such a reference. If you have one, please let me know.
Plus, are there any biases observed using this new kit?
Using the new illumina small RNA sample prep kit v1.5, do we expect an additional "A" right after the miRNA sequence?
Here is the original 3' RNA Adaptor sequence and result library and I added the red colored "A":
3' RNA Adapter
5' P-AUCGUAUGCCGUCUUCUGCUUGUidT
Result Library:
5' AATGATACGGCGACCACCGA CAGGTTCAGAGTTCTACAGT CCGACGATC (N) ATCGTATGCCGTCTTCTGCTT G 3'
3' TTACTATGCCGCTGGTGGCT GTCCAAGTCTCAAGATGTCA GGCTGCTAG (N) TAGCATACGGCAGAAGACGAA C 5'
I suspect that illumina improved the new kit using pre–5',5'-adenylated adaptor, so the adaptor ligation can be performed in absence of ATP. However, I am not able to find such a reference. If you have one, please let me know.
Plus, are there any biases observed using this new kit?
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