Could you tell me why there are multiple peaks in the 16S V3-V4 tapestation data?

Question

[remi]

Hi researchers.

I would like to know why my results of 1st PCR and 2nd PCR differ from illumina manual. https://support.illumina.com/documents/documentation/chemistry_documentation/16s/16s-metagenomic-library-prep-guide-15044223-b.pdf

I ordered the forward and reverse primers listed in the URL from the oligomer design company.

• 16S Amplicon PCR Forward Primer = 5' TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGCCTACGGGNGGCWGCAG
• 16S Amplicon PCR Reverse Primer = 5' GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAGGACTACHVGGGTATCTAATCC

I think there may be a problem with the primers.

Thank you for your answer.