Genomics technologies, in particular high-throughput sequencing and bioinformatics, have revolutionised modern biology and genetics. As a result, building capability in both areas is a critical strategic priority for the Institute, necessary to underpin the high quality research described in both Pirbright ISPs, Host and Virus. To achieve this, we have unified the CCG-related bioinformatics and high-throughput sequencing activities. A coordinated approach will enable us to get the best from both worlds: sequencing protocols that provide the most to subsequent bioinformatics, and data analysis workflows optimised for each protocol.
We provide a unique sequencing solution within high containment, with world class facilities. As work at the institute involves a number of SAPO4 viral agents unable to leave containment, this arrangement offers us the invaluable and unprecedented opportunity to perform cutting edge research on these agents in situ.
At the moment the facilities are mostly built around the Illumina ‘sequence by synthesis’ technology platform, but we are also exploring novel technologies providing longer reads, such as the Oxford Nanopore MinION.
We have several Illumina sequencer machines, each tailored to different output scales and specifications. They provide established workflows for genetics, transcriptomics and epigenetics, as per the Institute’s research requirements specified in the ISPs. Our DNA- and RNA-sequencing capabilities allow us to deploy core workflows that include whole genome sequencing, targeted amplicon, RNA and PCR-free applications. We collaborate closely with others at the Institute in the provision of training and resources for researchers to learn and perform library preparation in a dedicated ‘clean’ environment within the SAPO4 envelope.
As part of our key goals we have automated core sequencing pipelines using a Hamilton NGS Star, in order to standardise high quality sequencing library production. Dedicated staff within the unit have extensive experience in a wide range of protocols (including CHIP-seq, miRNA-seq and ribosome profiling) and can develop bespoke processes that cannot be supported by established high throughput sequencing pipelines. This is an important part of our strategy as we seek to maximise the sequencing potential across the diverse and important historical sample archives held within the Institute.
Another strength in our strategy is the emphasis on embracing novel and emerging genomics methodologies, ensuring science at Pirbright remains at the cutting edge of animal virology science. In line with this we are working to establish a strong single cell capability within the Institute, and have recently procured a Dolomite Bio micro-encapsulator. This new equipment will enable the deployment of droplet-based protocols, including drop-seq, immunological profiling and production of droplets for FACs sorted libraries.
The CCG-funded bioinformatics activities within the unit/group focus on the development and delivery of robust and highly sensitive analysis pipelines for the de-novo assembly of DNA/RNA viruses, RNA-sequencing analysis and tools aimed at the evolutionary analysis of viral/host sequences. These novel in-house pipelines have already been applied to a number of Pirbright projects including the characterisation of FMDV quasi-species in buffalo, the genomic analysis of vector species and the determination of biomarkers in Marek’s disease virus infection.
We are also committed to developing specialist tools, such as variant callers specialised for viral data, protocols for the high precision clustering of immune molecules and integrative tools to analyse data deriving from a combination of protocols and conditions.
The group also run Pirbright’s high performance computing cluster. With ~1000 hyper-threaded CPU cores, 4TB RAM and 0.5 PB of storage, it provides the group and all quantitative scientists at the Institute with a modern cutting edge high speed data analysis capability.
Sequencers within high containment include an Illumina MiSeq, NextSeq-550, iSeq-100 and Oxford Nanopore MinION. We have integrated these systems in with lab-based automation with a Hamilton ML-STAR. For single cell investigations and other droplet-based applications we have a Dolomite Bio microencapsulator.
Bioinformatics capabilities include the high performance computing cluster.