Dear Sequencing gurus:
I would like to prepare a sequencing library that includes an upstream sequence (for the purposes of performing template suppression PCR on the library). Here is the structure of the envisioned library:
TS Sequence - P5 TruSeq - LIBRARY SEQUENCE - P7 TruSeq - TS Sequence
The TS sequence would ideally be 27 bases long.
Do you think that would interfere with cluster generation? Anyone have experience putting sequence upstream of P5/P7? I can always cleave it off with a restriction enzyme, but I'd like to avoid that will reduce library complexity.
Thanks in advance for your comments!
I would like to prepare a sequencing library that includes an upstream sequence (for the purposes of performing template suppression PCR on the library). Here is the structure of the envisioned library:
TS Sequence - P5 TruSeq - LIBRARY SEQUENCE - P7 TruSeq - TS Sequence
The TS sequence would ideally be 27 bases long.
Do you think that would interfere with cluster generation? Anyone have experience putting sequence upstream of P5/P7? I can always cleave it off with a restriction enzyme, but I'd like to avoid that will reduce library complexity.
Thanks in advance for your comments!
Comment