I do not know if it is possible to analyze V3-V4 regions from 16S if I have only 150bp-reads (amplicon V3-V4 had 500bp aprox.), so there is no overlapping between reads F and R. With DADA2 and vsearch I get an error, and only Deblur allowed me to do the denoise of the reads, but I am not sure if I am doing well. Is it considered single-end instead of paired end as there is no overlapping between reads F and R? Now we are performing the sequencing with 301 cycles, instead of 151.
Thank you in advance!!