The molecular laboratories at RBGE are well equipped with facilities for molecular biodiversity research, from DNA extraction through to library prep for Next Generation Sequencing.
DNA Extraction Lab: Equipped for manual and automated DNA extraction: Qiagen TissueLyser, Fastprep-24 , thermomixers, centrifuges, vented benching, fume hood. This lab can be used for high molecular weight DNA extractions from fresh samples, but can also be converted for working with highly degraded DNA samples, with a set of dedicated low-copy DNA extraction equipment.
Pre-PCR Lab: Clean lab space dedicated to Pre-amplification work with DNA, including DNA banking and PCR set-up.
Post-PCR Lab: Clean lab space dedicated to Post-amplification work with DNA, including gel imaging and amplicon pooling.
Next Generation Sequencing Lab: DNA and library quality check equipment, including Agilent Bioanalyser, Tapestation 2200 and Femtopulse, DeNovix Spectrophotometer/Fluorometer, Qubit, Diagenode BioRuptor for DNA fragmentation, a thermomixer and various thermocyclers, including a BioRad CFX96 Real-Time System. An Oxford Nanopore Technologies MinION with a flongle adaptor is also available; RBGE now facilitates Oxford Nanopore Technologies’ amplicon sequencing in addition to traditional Sanger sequencing, with a highly cost-efficient method that allows simultaneous sequencing of multiple loci for multiple accessions.
Gel/RNA lab: Dedicated bench-space and equipment for extracting and working with RNA. This lab also contains gel-running apparatus, with a Syngene gel analysis system.
Secure access lab: External laboratory space for restricted-access molecular work, including foreign soils and plant pathogens.
Bioinformatics suite: Computers loaded with sequence editing and analysis programmes, including Sequencher and Geneious, and an analysis server.
Historic DNA lab: Low-copy DNA extraction and library prep lab with separate room for sample powdering. This lab is dedicated to extractions from herbarium material and is physically isolated from any post-PCR area. Contamination from environmental DNA and PCR products is kept to a minimum by ensuring one-way workflows, the use of PPE, a strict daily cleaning routine, and UV-decontamination of reagents and consumables. The lab is equipped with HEPA-filtered PCR and biological cabinets for sample and reagent handling in a cleanroom-like environment.