Whole Exome Sequencing

Service Description

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 NovaSeq X Plus.

Sample Requirements:

• DNA Quality:
• DNA Amount: 200ng (it is possible to generate libraries from much less DNA, however this will result in reduced library complexity (ie fewer unique library molecules after PCR) and as a result after removing PCR duplicates during analysis, desired X coverage may not be possible.
• Volume: >10ul
• 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.

Analytical Goals:

• Germline and somatic variant calling
• Copy Number Analysis (can also be done with low pass whole genome sequencing)

Other Information:

• Twist Exome Panel (bed file)
• Sequencing with paired-end 150bp reads

Recommended Bioinformatics Pipeline: Alignment and Variant Calling pipeline