Fragment analysis is run on our ABI 3130XL using a fluorescence-based detection system. The use of six fluorescent dyes (one color is reserved for size standards) allows multiplexing of fragments, increasing throughput and lowering costs. A size standard is included in every lane, eliminating lane to lane variability. We accept samples in 96 well plates or 8-tube strip tubes, and supply the LIZ 600 or ROX 625 size standard. To determine the proper concentrations of PCR product, we recommend you submit a small run of samples of varying dilution before starting a large project. Please see "Submitting a Sample" below for instructions on how to submit fragment analysis samples. For more information, please contact the Core Facility.
Supported Fluorescent Labels
The ABI 3130, using Dye Set G5, will detect fragments labelled with the fluorescent dyes FAM (blue), VIC (green), NED (yellow) and PET (red). The fluorescent dye HEX may be used in place of VIC, and TAMARA may be used in place of NED.
Commonly the LIZ 600 or ROX 625 standards are used, allowing for size determinations from 50 - 625 base pairs.
Samples must be submitted in 96-well plates or 8-tube strips, and organized in the 8 well format - A1–H1, A2-H2, etc. Please use V-bottom plates-flat-bottom plates are difficult to pipette from. Securely seal plates and strip tubes with caps (recommended) or sealing film. Label the plate on the side. If you have multiplexed samples, we recommend a small trial run to determine the optimal conditions.
Download the request form here. Fill in all the information, making sure the plate name on the plate is the same as the plate name on the form. Sample names cannot have spaces, periods, or Greek symbols-the software won’t accept them. Hyphens and underscores are acceptable. Email the completed form to email@example.com. Samples can be dropped off at Howell Science Complex, N203-please call ahead (328-2607) to make sure someone is available to receive your samples.