Publication details

A mcrA gene sequencing and analysis protocol for the Illumina MiniSeq sequencer



Year of publication 2019
Type Conference abstract
Description In the last decades methanogenic archaea have been increasingly investigated with regard to their impact on the environment, technological applications or role in human health. Today, high-throughput sequencing of the functional mcrA gene is routinely used to assess methanogenic archaea diversity in environmental samples. Compared to the 16S rRNA gene, the mcrA gene provides higher sensitivity in analysis of methanogens phylogeny. In our study we propose sequencing approach for high-throughput sequencing of the functional mcrA gene from complex communities on the MiniSeq sequencer by using sequencing primers that enable dual-index barcoding. A mock community was sequenced alongside the environmental, biotechnological and human samples in three different sequencing runs using classical mlas/mcrA-rev primer set and 16S rRNA primers amplifying V4 region. In both mcrA sequencing runs we were able to recapture a realistic composition of the mock community, and find similar differences in the environmental, biotechnological and human samples. However, in 16S rRNA sequencing run we registered slight differences in alpha and beta diversity within the samples. Our results demonstrate that the MiniSeq platform can produce similar quantities of mcrA reads compared to the MiSeq platform and thus represents cost-effective option in performing high-throughput mcrA gene sequencing.

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