I have a requirement to introduce variable-length non-specific regions into my library as every molecule contains T3/T7 universal tails and thus would result in cluster-recognition failure as the first 20/21bp is identical. For this reason I need to do the initial library amplification using custom primers before moving on to the use the NEBNext Index primers for paired-end sequencing on a MiSeq.
Essentially the primer structure would be:
Illumina sequence-N(1-4)-T3/T7
I've not carried out a MiSeq run before so i'm mostly looking for reassurance and a straight answer as to what sequences I would need to include before I could go ahead and use the index primers and introduce the P5/P7 sequences.
What would they be?
- Julia
Essentially the primer structure would be:
Illumina sequence-N(1-4)-T3/T7
I've not carried out a MiSeq run before so i'm mostly looking for reassurance and a straight answer as to what sequences I would need to include before I could go ahead and use the index primers and introduce the P5/P7 sequences.
What would they be?
- Julia