The MRF DNA sequencing facility has been in operation since 1995. This facility has been highly successful and faces continually increasing demand. Our philosophy is to provide high quality data and expertise as inexpensively as possible. Please follow our Guidelines for the best quality sequencing data.

Users submit template and, where relevant, custom primer in separate tubes according to the following guidelines. Templates should be purified by methods described below. The MRF staff performs the sequencing reaction, generates sequence data, performs data analysis and delivers data as indicated below. For Full Service sequencing, DNA samples are sequenced using ABI BigDye Terminator chemistry on an ABI 3130xl Genetic Analyzer.

 

TEMPLATE
SS DNA
DS DNA
PCR PRODUCT
Concentration
0.2 µg/µl
0.2 µg/µl
10 ng/µl
solvent
Water (not TE)
Volume (per reaction)
5 µl*
5 µl*
10 µl*
Submit sample in labeled...
1.5 ml microfuge tube
PRIMER
SS DNA
DS DNA
PCR PRODUCT
Concentration
50 ng/ul or 9 pmole/µl
Volume (per reaction)
5 µl*
Submit Sample in a labeled
1.5 microfuge tube

*Minimum volumes are listed. If requesting more than 1 sequence for a primer or template, add an additional 2 µl for each additional sequence requested.

Turnaround Time: Maximum 20-48hr.

Policy: OUR POLICY IS TO RE-RUN FULL SERVICE SEQUENCING REACTIONS FREE OF CHARGE when we feel we can improve run quality. This policy only applies if our specifications are followed, and if complete template and primer information is provided on the on-line sequencing request form.

Charges: Our charges for each project will vary but will reflect the current prices shown in our Price List.

Facility Supplied Primers:(Free of charge) M13 F, M13 R, SP6 promoter, T7 promoter, T7 terminator, T3 promoter, BGHrev, GL1, GL2, RV3, RV4, pGEX3, pGEX5.

Editing: The MRF can edit your sequence for you. This consists of, correcting the analyzed sequence called by the sequencer. We scan the entire sequence for ambiguities or miscalls, and, where possible, manually correct these errors. For viewing and editing your own data, download a free or shareware version of Chromas software for a PC or free Editview 1.0.1 and PC-Mac softwares for Mac. Mac users have to first convert sequence data files into Mac version using PC-Mac program, then open files using Editview.

NEW: Use a freeware or shreware version of ChromeView for PC and Mac or use online version of ChromeWeb to open electropherogram files.

Data Delivery: Notice is sent via e-mail to the contact person listed on the order form when the sequences are ready. Both electropherogram files(.ab1) and text files (.seq) can be viewed and downloaded from the site using your order number.

Large DNA Templates: Large templates including Cosmids, Lambda, BAC's and chromosomal DNA can be sequenced using a specialized reaction. Please inquire about the special requirements to accomplish sequencing from these templates.

The MRF will "walk" your gene of interest including primer design, primer synthesis and purification, automated sequencing, sequence assembly and consultation.


Primer Design: The MRF will design primers to the highest compatibility with your DNA template. Primers will be synthesized and purified by the MRF. Extra primer will be provided to you accompanied by a printout of the primer sequence, concentration, and other data upon project completion.

When ordering, keep in mind that a good sequencing run with one standard primer reads about 700 to 800 bases, but that custom primers often give variable results. Please be aware that there are many steps to assuring the quality of your data, so the project may take some time.
For any questions please contact the DNA Sequencing lab by calling (973) - 972 - 2625 or by emailing mrfadm@njms.rutgers.edu