Tag: molecular labPage 1 of 2

Polytrichum baits: cutting cleaned mosses

Following on from the rather unpredictable results we obtained from fragmenting duplicate aliquots of CTAB-extracted Polytrichum DNA in the Bioruptor, Isuru cleaned aliquots of IK31 and IK53 using…

Hybrid capture from degraded DNA: the mysterious case of the vanishing libraries

When we’re working out a protocol or troubleshooting, we spend a lot of time quantifying small quantities of fluids, looking at DNA concentrations on the DeNovix, running tapes…

Polytrichum baits: Cutting mosses

We started our lab work on the Polytrichum hybrid baits project on the 1st of October, by normalising some CTAB-extracted DNA with 0.1X TE to 55 µL of…

Generating a phylogeny of Polytrichum using hybrid baits

The current Next Gen Sequencing lab project at the Botanics involves looking at the phylogeny of Polytrichum section Polytrichum, using hybrid capture. The work will form part of…

Hybrid capture from degraded DNA: bead-cleaning away the adaptor peaks

After testing bead-to-sample ratios of 30:50, 35:50, 40:50, 50:50 and 60:50 on a Thermo Scientific™ GeneRuler™ 50 bp DNA Ladder, using Beckman Coulter AMPure XP beads, we focused…

Hybrid capture from degraded DNA: testing bead cleaning on a 50bp ladder

When we made our Begonia libraries, working with (in some cases) relatively small quantities of very degraded DNA, we should have diluted the adaptors. We didn’t. Consequently, we…

Hybrid capture from degraded DNA: quality metrics for duplicate extractions

For one of the taxa in our study set, Begonia scottii (living collection no. 20170076), we made a few replicate DNA extractions using Qiagen DNeasy plant mini-kits, and…

Hybrid capture from degraded DNA: bead cleaning degraded Begonia DNAs

The set of DNA extractions from one of our test plants (#8, Begonia stictopoda RBGE accession 20170115) contained rather low concentrations of DNA. We decided against preparing NGS…

How much liverwort do you need to get 50 μg of DNA?

There’s an exciting project, The 10KP (10,000 Plants) Genome Sequencing Project, that aims to sequence and characterize representative genomes from every major clade of embryophytes, green algae, and…

Alternative Methods for Collecting Plant Material for Future DNA Extraction – Part I

Quality and quantity of DNA recovered from stored plant material is becoming more important as DNA sequencing technologies change from the relatively simple Sanger sequencing, to next generation…

Hybrid capture from degraded DNA: test Begonia sample quality

Keen to see the effects of different specimen preservation techniques on DNA quantity and quality, we have assessed extractions of DNA from nine Begonia accessions x seven preservation…

Hybrid capture from degraded DNA: Squashing Begonia

In order to look at the effects of herbarium preservation methods on DNA quality, Hannah Wilson and Mark Hughes took a trip down to our research glasshouses, and…

Hybrid capture from degraded DNA: stockpiling Begonia DNA for protocol testing

When working out new protocols, it’s very useful to have a big stash of DNA to test them on. Just now, in a collaborative project with Dr Michelle…

Untangling asetate Weissia species in the UK

Dr Des Callaghan spends rather a lot of his time chasing after rare things. He’s an environmental consultant with many strings to his bow, but a particular specialisation…

Nuclear internal transcribed spacer (ITS) primers

There are lots of different primers that sit in slightly different parts of the ITS locus, some forward and some reverse. There’s a very well-worn printout stored on…

Bryological holiday jobs

The Science building at the Botanics closes down between Christmas and New Year, so any last bits of work for the year have to be packed up and…

DNA Sequencing Natural History Specimens Using New Sequencing Platforms and Protocols: a 1-day meeting at RBGE 11/07/2017

Rapid developments in high-throughput sequencing platforms are providing a step change in the recoverability of DNA sequence data from natural history collections. Short-read massively parallel sequencers are intrinsically…

Testing extractions – comparing DNA on agarose gels

Looking at the capture plates from the two DNA extraction protocols that were tested on our QIAcube, it was fairly obvious that a lot more plant fragments and…

Describing your DNA

One of the amazing things about the polymerase chain reaction, PCR, is how little starting DNA is needed, with an exponential increase in the number of copies of…

Letting the robot do its job

Having got together two plates of tubes with little bits of plant and lichen tissue in them, and pulverised them with tungsten beads in a TissueLyser for a…