Hi,
I have a plasmid library that looks like this:
uniq-seq1___45bp__sequence-of-interest___45bp__uniq-seq2
The 45bp sequences are identical and oriented as inverted repeats. I'd like to sequence the middle region and maintain orientation (so using primers in the unique sequences that flank the repeats (green/purple)). Will this be possible on a MiSeq, or will I run into problems in cluster generation or sequencing because of the inverted repeats? I can PCR this amplicon without much trouble, so I'm not too worried about generating the library for sequencing.
The repeats are 67% A/T if it matters.
thanks for any help you can provide
-Lucas
I have a plasmid library that looks like this:
uniq-seq1___45bp__sequence-of-interest___45bp__uniq-seq2
The 45bp sequences are identical and oriented as inverted repeats. I'd like to sequence the middle region and maintain orientation (so using primers in the unique sequences that flank the repeats (green/purple)). Will this be possible on a MiSeq, or will I run into problems in cluster generation or sequencing because of the inverted repeats? I can PCR this amplicon without much trouble, so I'm not too worried about generating the library for sequencing.
The repeats are 67% A/T if it matters.
thanks for any help you can provide
-Lucas
Comment