Comment on Bohmann et al. Strategies for sample labelling and library
preparation in DNA metabarcoding studies
Abstract
DNA metabarcoding necessitates labelling amplicons in order to connect
sequencing reads with samples, but labelling protocols may cause errors
where indexes are incorrectly assembled during PCR due to tag-jumping. A
recent paper by Bohmann et al (2021) reviews the main labelling methods
and point out that library building using PCR’s on tagged amplicons may
be particularly problematic. Due to unforeseen problems in two
sequencing projects, we had to use a second PCR on tagged amplicons to
salvage two large data sets. This test showed that the problems with
tag-jumping errors were acceptable and could be accounted for during
analysis, if handled properly when designing the indexing strategy.