Hi everyone, does anybody have any ideas for things I can try to get rid of this non-specific amplification? These products are for Illumina sequencing - I used barcoded V4 16S primers below:

F: AATGATACGGCGACCACCGAGATCTACACATGCCTAATATGGATAATTGTGTGYCAGCMGCCGCGGTAA

R: CAAGCAGAAGACGGCATACGAGATATGCCTAAAGTCAGCCAGCCGGACTACNVGGGTWTCTAAT

The PCR conditions I used were:

94 °C - 3 min

94 °C - 45 s

50 °C - 60 s

72 °C - 90 s

x35

72 °C

10 min

How problematic will this be for the sequencing? I'm thinking increase the annealing temp, but to what? Thank you so much for any help!