Next generation DNA sequencing allows study of the genome, transcriptome and epigenome of organisms as well as using more targeted experiments to examine how our genome is structured and used.
The MiSeq uses Illumina SBS chemistry to sequence pre-prepared sample libraries giving millions of individual fragment sequences. The sequences produced will give information determined by your experimental design. For example, this could be genomic sequence to allow assembly of a genome or transcriptome sequence to provide information on gene expression. Output data is supplied as FASTQ files.
The MiSeq can be used for genome sequencing, transcriptome analysis, amplicon sequencing and metagenomic studies as well as many more specialised applications. The MiSeq offers longer sequencing reads of up to 300bp as well as reduced running time for fast data turnaround.
How does it work?
A sample library is made consisting of DNA fragments with known DNA sequences attached to either end (adaptors). This sample is loaded on to the MiSeq and the adaptor sequences along with on board chemistry (with reagents delivered via microfluidics across a glass flowcell) allow the library fragments to anneal to the flowcell. Once the fragments have annealed a process called cluster generation is carried out which amplifies the DNA fragments to enable to visualisation of these clusters during the sequencing chemistry cycles. Each cycle of chemistry allows one base of each DNA fragment to be identified and this is repeated to collect the sequence information of the DNA fragments.
Pre-made libraries or library pools require QC information on library size and concentration.
Dr Ian Hancox, 024 76 150 380
email i dot hancox at warwick dot ac dot uk.
Mrs Jeanette Selby, 024 76 5 75172
email j dot m dot selby at warwick dot ac dot uk.
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