21 07, 2017

DAP-Seq for higher-throughput transcription factor analysis

By | July 21st, 2017|Categories: Methods and applications, Next-generation sequencing|0 Comments

The Ecker lab at the Salk Institute published a method for very high-throuphut transcription factor analysis in Nature Protocols yesterday: Mapping genome-wide transcription-factor binding sites using DAP-seq. The method is elegantly simple in its concept; PCR-free libraries from native gDNA are hybridised with affinity-tagged in vitro-expressed transcription factors, […]

12 07, 2017

ctDNA analysis from 250pg

By | July 12th, 2017|Categories: Methods and applications, Next-generation sequencing|2 Comments

A recent paper in Nature Scientific Reports from Maurice Jansen’s group at the Erasmus Medical Centre in Rotterdam presents ctDNA analysis by targeted NGS from just 250pg of input cfDNA . This is a very small amount of material, and although the authors do discuss […]

11 05, 2017

Update on @illumina index-swapping: better libraries

By | May 11th, 2017|Categories: Methods and applications, Next-generation sequencing|Tags: |3 Comments

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 […]

5 05, 2017

Inflection points for @nanopore sequencing

By | May 5th, 2017|Categories: Methods and applications, Nanopore sequencing, Next-generation sequencing|6 Comments

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 […]

24 04, 2017

Update on @illumina index-swapping: better barcode design

By | April 24th, 2017|Categories: "Experimental design controls etc", Core facilities, Methods and applications, Next-generation sequencing|Tags: , , |2 Comments

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! […]

21 04, 2017

Update on @illumina index-swapping

By | April 21st, 2017|Categories: "Experimental design controls etc", Core facilities, Design, Methods and applications, Next-generation sequencing|0 Comments

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 […]

7 04, 2017

@qiagene GeneReader data published

By | April 7th, 2017|Categories: I am not a clinician, Next-generation sequencing|Tags: , |0 Comments

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 […]

28 03, 2017

What does RNA-Seq mean to you

By | March 28th, 2017|Categories: Core facilities, Methods and applications, Next-generation sequencing|1 Comment

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 […]

28 03, 2017

HMW DNA extraction for long-reads on @nanopore or @10XGenomics

By | March 28th, 2017|Categories: 10X Genomics, Methods and applications, Next-generation sequencing|8 Comments

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 […]

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