Index-swapping appears to be driven by excess adapter/primer (Illumina whitepaper). The take-home messages are 1) to use UDIs (unique dual-indexes), and 2) clean-up your libraries to remove left-over adapter/primer. Later in this post I’m going to work through one solution for getting rid of any remaining adapter/primer in an NGS library; but […]
When will nanopore sequencing push short-reads (i.e. Illumina) off their pedestal? According to Clive PromethION is the Illumina killer…but this same conversation was going on many times at London Calling. I wanted to highlight two areas that might be about to flip to ONTs advantage […]
Last week I followed up on the index-swapping issue after Illumina released their white paper and also covered what Ethan Linck at The Molecular Ecologist had posted about the Sinha et al BioRxiv paper. In that post I said I’d write a follow-up post about index design over the weekend – here it is! […]
Illumina released a white paper this week describing their efforts to understand the index-swapping issues with the ExAmp chemistry. This story has been covered many times and got lots of Tweets in the days since the BioRxiv paper from Stanford was uploaded. Megan Molteni at Wired wrote a lengthy […]
The Qiagen GeneReader is an NGS platform developed for the clinic and aims to deliver a sample-to-answer solution. I covered the “launch” of the instrument at the end of 2016 and summarised some of the details but there was no public data to take a […]
This post is a follow on from my ABRF2017 presentation this morning. Slides are available on SlideShare. RNA-Seq is the most widely published NGS method, but what do researchers mean by the term “RNA-Seq”? Virtually none of them mean sequencing of RNA. Most are talking […]
Long-reads require long DNA molecules which require high molecular weight (and undamaged) DNA. Extraction with your standard column is unlikely to be optimal. I’m starting to look at the best methods for HMW DNA extraction for long-read applications as my lab is supporting a couple of […]
Anyone interested in single-cell RNA-Seq analysis is likely to be interested in a paper published today by the Sanger in Nature Methods: Kiselev et al. (2017) SC3: consensus clustering of single-cell RNA-seq data. The new single-cell clustering tool performs better than 5 other methods: tSNE + k-means […]
Confession: this post may not actually make your 10X Genomics experiments ten times cheaper…but implement Demuxlet in your workflow and with the right sort of experiment you might just get there! Single-cell RNA-Seq might just have got a whole lot cheaper, thanks to a method […]
In 2013 researchers from Oxford’s Molecular Diagnostics Centre, NIHR BRC, WTCHG and Department of Oncology led by Anna Schuh and Jenny Taylor sequenced 351 lung, melanoma and colorectal tumours from patients using an NGS 46-gene hotspot cancer panel. Results were delivered with a median turnaround of 7 days […]
