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We are planning to run human exomes on Illumina's Nextseq and I just wanted to confirm that I got some details right.
We will use the TruSight One and Rapid Capture Kits and I have a small question about dual indexing.
We aim to pool 12 whole exomes (Nextera Rapid Capture) or 36 clinical exomes (TruSight One) on one Nextseq 500 High Output 2X150 run.
So for the first case, I would use single index sequencing (12x index 1 primer = N701- N712) and for the second case I would use dual indexing (12 i7 x 3 i5).
My understanding is, that in both cases, each reaction needs to have a i7 and a i5 primer, but in single indexing the i5 is just not read. Is that correct or is the i5 optional for single indexing (I doubt it)? So the only difference in setting up the run would be to add the BP13 (index 2 primer mix) to the reagent cartridge and choose dual indexing in the run parameters in Basespace.
thx
We will use the TruSight One and Rapid Capture Kits and I have a small question about dual indexing.
We aim to pool 12 whole exomes (Nextera Rapid Capture) or 36 clinical exomes (TruSight One) on one Nextseq 500 High Output 2X150 run.
So for the first case, I would use single index sequencing (12x index 1 primer = N701- N712) and for the second case I would use dual indexing (12 i7 x 3 i5).
My understanding is, that in both cases, each reaction needs to have a i7 and a i5 primer, but in single indexing the i5 is just not read. Is that correct or is the i5 optional for single indexing (I doubt it)? So the only difference in setting up the run would be to add the BP13 (index 2 primer mix) to the reagent cartridge and choose dual indexing in the run parameters in Basespace.
thx