The whole exome sequencing method (WES) begins with the commercially available Library Preparation EF Kit 2.0 from Twist BioSciences. Resulting libraries are then pooled and exome containing sequences captured using Twist Target Enrichment Fast Hybridization reagents. Enrichment probes cover a 37.7 Mb target region in the human genome. The uniform coverage of the target regions (98%) with >30x coverage provides optimal performance for sequecing of exomic regions.
Unique dual indexes are available for multiplexing of up to 96 samples. Genomic DNA input is 50 ng.
Core staff are available to assist with experimental design and to answer questions related to next-generation sequencing technology. Arrangements for a meeting to discuss new or existing projects can be made by sending an email to Nathan Bivens, Genomics Core director.
All projects must be initiated with a quote before samples are submitted. Send a quote request to the general core email mugenomicscore@missouri.edu.
The Genomic Calculator may be used to estimate the amount of sequence required for a project.
General Considerations:
- Submit samples only after paperwork has been completed.
- Clearly label tubes. Names must match those given on the project information form.
- Low binding microcentrifuge tubes (0.5 or 1.5 mL) are preferred for storing high quality nucleic acids.
Submission Requirements:
| Input Range | Submission Concentration |
Recommended Buffer |
Special Instructions |
| 50 ng | 25 - 100 ng/μl genomic DNA | 10mM Tris (pH 8.5) | An image of 100 ng of genomic DNA following agarose gel electrophoresis and staining will be provided to demonstrate gDNA integrity. |
| Sequence Depth/Coverage* | Read Type | Index Type | Read Length | ||
| 100x coverage | Paired end | Unique dual index | 100 bases | ||
* coverage recommended by MUGTC for mammalian genome; increased coverage may be desired for some studies