Information on submitting samples for Genotyping-by-Sequencing (GBS)
GBS services are only for OSU internal projects. Contact Brett Tyler if you have questions about this policy.
Samples provided in plate format.
Two files to upload with your order: Sample ID sheet (as described under Sample Quantity below) and gel image (as described under Sample Quality below).
GBS: The quantity needed for a GBS experiment is 250ng/sample normalized across all samples to the same concentration (no less than 20ng/ul and no more than 100 ng/ul). Sample ID sheet includes the Sample name associated with plate location, the concentration and volume submitted and any additional pertinent information about the sample.
Determining adapter concentration: If you are using a unique organism/enzyme combination and the adapter titration value has not been determined, then an optimization step must precede the GBS experiment. One sample at 2 micrograms (ug) of high quality gDNA is needed for the Titration / Optimization experiment. In most cases, an adapter optimization experiment is not necessary.
We recommend generating a gel image showing all samples uncut and a subset of these samples (10%) which have undergone restriction digest using an enzyme which is NOT methylation sensitive (i.e. EcoRI, HindIII). A 1% agarose gel is often sufficient. The Λ HindIII digest (ladder) is required.
DNA quality is critical for success in a GBS experiment. Uncut genomic DNA should have a tight band greater than 10Kb while restricted samples should exhibit a smear running from 0.2-10KB. We do not complete a test digestion before completing library preparation.