Direct PCR meets high-throughput sequencing – metabarcoding of chironomid communities without DNA extraction
- Aquatic emergent insect communities form an important link between aquatic and terrestrial
ecosystems, yet studying them is costly and time-consuming as they are usually
diverse and superabundant. Metabarcoding is a valuable tool to investigate arthropod
community compositions, however high-throughput applications, such as for biomonitoring,
require cost-effective and user-friendly procedures. To investigate if the time-consuming
and labour-intensive DNA extraction step can be omitted in metabarcoding, we
studied the difference in detection rates and individual read abundance using standard
DNA extraction versus direct PCR protocols. Metabarcoding with and without DNA extraction
was performed with artificially created communities of known composition as
well as on natural communities both of the dipteran family Chironomidae to compare
detection rates, individual read abundances and presence-absence community composition.
We found that the novel approach of direct PCR metabarcoding presented here
did not alter detection rates and had a minor effect on individual read abundances in
artificially created communities. Furthermore, presence-absence community compositions
of natural chironomid communities were highly comparable using both approaches.
In conclusion, we showed that direct PCR protocols can be applied in chironomid
metabarcoding approaches, with possible application for a wider range of arthropod
taxa, enabling us to study communities more efficiently in the future.