Whole Exome Sequencing (WES)
Method
Purified genomic DNA is fragmented using a Covaris R230 AFA instrument. Whole genome libraries are prepared using IDT XGen DNAseq reagents on a Beckman Coulter Biomek i7 liquid handling platform. Finished libraries are pooled (typically 8-plex) and exome hybridization capture enrichment is performed using Twist Biosciences probes and reagents. Sequencing is performed with paired-end 150bp reads to specified depth of coverage on an Illumina NovaSeq6000.
Pricing
QC + library prep + sequencing
(50X Coverage)
DFCI/BWH: $450
External: $500
Alignment, Variant Calling, Filtering:
+$20/sample
Turnaround
3 weeks
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DNA QC
DNA is assayed by qubit to determine concentration
Analytical Goals
Variant Calling
Sample Requirements
Sample: >200ng of purified gDNA (we can work with less, 50ng min.)
Buffer: EB (10mM Tris-Cl, pH 8.5) or MolBio. grade nuclease free water. Buffers containing EDTA should be avoided because it may inhibit enzymatic reactions.
Volume: >10ul (typically 20-30ul)
Container: 1.5mL Eppendorf tubes (preferred)