Tag: PacBio

On Friday, Molecular Ecology Resources put online Christian Wurzbacher‘s latest paper, of which I am also a coauthor. The paper presents three sets of general primers that allow for amplification of the complete ribosomal operon from the ribosomal tandem repeats, covering all the ribosomal markers (ETS, SSU, ITS1, 5.8S, ITS2, LSU, and IGS) (1). This paper is important because it introduces a technique to utilize third generation sequencing (PacBio and Nanopore) to generate high‐quality reference data (equivalent or better than Sanger sequencing) in a high‐throughput manner. The paper shows that the quality of the Nanopore generated sequences was 99.85%, which is comparable with the 99.78% accuracy described for Sanger sequencing.

My main contribution to this paper is the consensus sequence generation script – Consension – which is available from my software page. Importantly, there are huge gaps in the reference databases we use for taxonomic classification and this method will facilitate the integration of reference data from all of the ribosomal markers. We hope that this work will stimulate large-scale generation of ribosomal reference data covering several marker genes, linking previously spread-out information together.

Reference

1. Wurzbacher C, Larsson E, Bengtsson-Palme J, Van den Wyngaert S, Svantesson S, Kristiansson E, Kagami M, Nilsson RH: Introducing ribosomal tandem repeat barcoding for fungi. Molecular Ecology Resources, Accepted article (2018). doi: 10.1111/1755-0998.12944 [Paper link]

As the 8th Next Generation Sequencing Congress in London is drawing to a close as I write this, I have a few reflections that might warrant sharing. The first thing that has been apparent this year compared to the two previous times I have visited the event (in 2012 and 2013) is that there was very little talk about where Illumina sequencing is heading next. Instead the discussion was about the applications of Illumina sequencing in the clinical setting; so apparently this is now so mainstream that we only expect slow progress towards longer reads. Apart from that, Illumina is a completed, mature technology. Instead, the flashlight is now pointing entirely towards long-read sequencing (PacBio, NanoPore) as the next big thing. However, the excitement around these technologies has also sort of faded compared to in 2013 when they were soon-to-arrive. Indeed, it seems like there’s not much to be excited about in the sequencing field at the moment, or at least Oxford Global (who are hosting the conference) has failed to get these technologies here.

What also strikes me is the vast amounts of talk about RNAseq of cancer cells. The scope of this event has narrowed dramatically in the past three years. Which makes me substantially less interested in returning next year. If there is not much to be excited about, and the focus is only on cancer sequencing – despite the human microbiota being a very hot topic at the moment – what is the reason for non-cancer researchers to come to the event? There will need to be a stark shift towards another direction of this event if the arrangers want it to remain a broad NGS event. Otherwise, they may just as well go all in and rename the event the Next Generation Sequencing of Cancer Congress. But I hope they choose to widen the scope again; conferences discussing technology as a foundation for a variety of applications are important meeting points and spawning grounds for novel ideas.