Next Generation Sequencing (NGS) and single cell experiments require careful planning and can be discussed with the facility and bioinformatics cores prior to commencement. NGS requires a sample to be converted to a sequencing ‘library’ before it can be sequenced. Sample libraries consist of fragmented sample DNA with the necessary sequencing adapters at both ends. These libraries can be mixed and sequenced together provided each has a unique barcode incorporated during preparation. The number of sample libraries which can be pooled together, and the choice of sequencer, depends on how much ‘coverage’ is required for each sample. The sequencing read length and strategy chosen (eg. 100 bp paired-end reads) will depend on the experimental aims.
The 10X Genomics Chromium system has a range of single cell applications including 3’ expression, 5’ expression and VDJ, ATAC and CNV. All 10X kits are now compatible with custom feature barcoding enabling Cite-seq and cell hashing experiments to be performed.
The qRT-PCR machines (Biorad CFX and ABI Quantstudio 7) are available for SAHMRI staff use and can be booked using the PPMS system.
The first step in the process is to discuss the project with the facility and obtain a quote. Where bioinformatics support is required other staff may be involved as necessary. To proceed with a job please return the quote with a purchase order number (external SAHMRI clients) or cost code.