Illumina Exome Sequencing
DNA is fragmented to 250bp using standard Covaris sonication. Fragmented DNA is then prepared as a standard Illumina gDNAlibrary using IntegenX reagents on the Apollo instrument, where the fragments are end repaired, A tailed, and adapter ligated. Then the samples are PCR amplified. Libraries are checked for quality and quantity, then pooled for capture 2-6 samples per pool (based on your request and your bioinformaticist’s advice). Each pool then undergoes exome capture (enrichment) using the Roche Nimblegen SeqCap EZ v3.0 according to the manufacturer’s protocols (Roche Nimblegen, Indianapolis, IN). The SeqCap EZ Human Exome Library v3.0 covers 64 Mb of coding exons and miRNAs. This service yields the exome-enriched libraries, but sequencing is an extra expense. Please consult with your bioinformaticist to determine appropriate parameters for sequencing, and required depth of sequencing.
|Platform||Illumina HiSeq 2500, HiSeq 4000, and NovaSeq.|
|Typical Cost||Click here to view internal pricing list!
Click here to view external pricing list!
|Typical Output||Illumina ready library|
|How to Request||Sample Submission: login through https://client-seqcore.brcf.med.umich.edu/ then choose ‘Illumina Sequencing’|
|Sample Requirements||1ug of good quality genomic DNA|
|Critical Information||We only stock the human kit. If you are looking for exome analysis of another species please contact us.|
|Additional Information||Due to limited space in the cores freezer's we can only keep samples for 6 months. If you would like your samples returned to you after sequencing please contact the core.
Due to size limitations on the HiSeq 4000, libraries with a average bp great than 580bp are not recommended to run on the HiSeq 4000.