Hi all,
I am new to deep sequencing and wish to receive your advice.
I am going to use NEBNext Ultra Directional RNA Library Prep kit with NEBNext® Multiplex Oligos for Illumina (Index Primers Set 1) and Miseq reagent kit v2.
I have several questions below
1) The index primer set 1 include a PCR universal primer and 12 index primers. I suppose dual-indexed library prep is impossible with this index primer set, am I right?
2) If so, I can only perform multiplexed single-read sequencing, NOT multiplexed paired-end sequencing as the latter requires dual-indexed libraries?
3) If multiplexed single-read sequencing is the only possibility, I should prepare libraries longer than 300bp and run 300 cycles?
Thank you very much in advance.
nvd
I am new to deep sequencing and wish to receive your advice.
I am going to use NEBNext Ultra Directional RNA Library Prep kit with NEBNext® Multiplex Oligos for Illumina (Index Primers Set 1) and Miseq reagent kit v2.
I have several questions below
1) The index primer set 1 include a PCR universal primer and 12 index primers. I suppose dual-indexed library prep is impossible with this index primer set, am I right?
2) If so, I can only perform multiplexed single-read sequencing, NOT multiplexed paired-end sequencing as the latter requires dual-indexed libraries?
3) If multiplexed single-read sequencing is the only possibility, I should prepare libraries longer than 300bp and run 300 cycles?
Thank you very much in advance.
nvd
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