i have tried 3 times to sequence 16S v3-V4 region, primer design is same like in earth microbiome project, reverse primer is exactly same , but forward is 338 f instead of 515f,
quality of reverse read is very poor, even we have included phix 10%
Earlier we did v4 region sequencing and it was perfect,..
totally confused, how same primer is perfect for v4 region but not for v3-v4, i have changed forward primer not reverse.
best
quality of reverse read is very poor, even we have included phix 10%
Earlier we did v4 region sequencing and it was perfect,..
totally confused, how same primer is perfect for v4 region but not for v3-v4, i have changed forward primer not reverse.
best
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