SOLS Shared Resource Facility

Overview of Services

SOLS Shared Resource Facility is run as a service. Laboratories may submit samples along with the sequencing request form. Sequencing reactions are usually run in our lab. However, we will run, at reduced cost, sequencing reactions — run in other laboratories. We will purify the reactions before running.  We maintain a stock of common plasmid primers accessible on the sequencing form. We use an Applied Biosystems 3730 capillary sequencer. Read lengths are routinely 800–900bp. Turnaround time is 24–48 hours. We can also perform plasmid or pcr purifications in conjunction with sequencing.

 Submitting Sequencing Samples

Purity and quantitation of the DNA template and primer dictate the quality of the sequencing results. The templates should be purified and quantified before submitting for sequencing. We can quantify templates for you, especially off campus customers; however this may slightly delay the return of your data.

Optimal amounts for templates and primers are as follows:

• Single stranded DNA 100-200 ng
• Double stranded DNA (plasmids) 150-200 ng
• PCR products 10-30 ng (10ng for templates <500bp; 30ng for >1kb)
• BAC or Cosmid DNA 500 ng

The amount of primer required for all of the above is 20 ng, except BAC and Cosmid which need 100 ng primer.

If you wish to set up reactions yourself, mix the template and primer at above amount and bring the volume to 6 ul with dd H2O. For off campus customers we recommend drying the sample prior to shipment to avoid possible loss, however properly sealed tubes can be successfully sent too. If you prefer that we set up reactions for you, submit sufficient template and primer in separate tubes.

Fragment analysis, which includes microsatellites, AFLP, or STR applications, are run on the 3730 sequencer. Dye-labeled samples are normally generated in respective laboratories and delivered to our lab for analysis. Contact us if you have questions on dye compatibility and before submitting samples.

Real time PCR in the SOLS SRF is run on an ABI7900HT thermocycler using a 384 well format. Samples are usually set up in respective laboratories. We provide training on the software and help in experimental setup, if required. Primers and probes can be designed using ABI Primer Express software available in the lab. Reservations are taken for three daily time slots.

The Agilent 2100 Bioanalyzer is a capillary electrophoresis unit capable of separating RNA, DNA or protein in lieu of gel electrophoresis. You may submit samples as a service request or you can be trained how to use the instrument. We routinely run RNA and DNA samples only, so please contact us before planning to submit any other samples. Chip types usually on hand are DNA1000, High Sensitivity DNA, RNA6000 Nano and RNA6000 Pico. Reservations are taken for daily use.

The Biotek HT1 Plate Reader scans multiple plate types in Absorbance, Fluorescence or Luminescence modes. Training is provided on the software and plate setup. The HT1 is equipped with gas control to allow anaerobic measurements or measurements under other environmental conditions. DNA and protein quantitation can also be done in a small volume 16 well plate. Reservations are taken for daily use.

Location and hours of operation

Hours: Monday - Friday, 7 AM - 4:30 PM

Contacts

Available Equipment and Resources