-Please send us a completed Request for Services form. This form can be found on the Web by clicking here or at http://www.umbi.umd.edu/cbr/core-facilities/dna-sequencing/images/cbrdna.pdf

Templates will not be sequenced without completion and submission of this form. (Users should keep a copy of this form for future reference.)

-A solution of template DNA or total nucleic acidin distilled water in a microfuge tube or microtiter plate, labeled by investigator's initials (on the tube cap) and sample ID and date of submission (both on the outside wall of the tube). Make sure there is no EDTA in the solution, as this reduces sequencing signal strength. A low amount of tris (e.g., 2-10 mM) is probably ok. The template can be plasmid DNA, phage DNA, or PCR fragments, either double- or single-stranded. PCR templates must be free of primer sequences. The samples submitted for sequencing should contain no radioactivity. Typically, DNA to be sequenced is not denaturated prior to thermal cycle sequencing. However, a few plasmid samples have required base denaturation or cleavage with restriction enzymes for satisfactory results.

-Appropriate sequencing primer in a microfuge tube. The primer concentration should be 0.8 pmole/ul. There should be 10ul of primer solution for each sample. M13-21 and M13REV primers are provided by the Facility.

M13REV CAG GAA ACA GCT ATG ACC
M13(-21) TGT AAA ACG ACG GCC AGT

-If you wish to get back the unused portion of your templates and/or primers, then you should submit these materials in a small, covered box labeled with your name and date of submission.

-One blank, IBM-formatted 3.5 diskette with investigator's initials or last name to hold any number of (uncorrected) sequences in text format OR one blank, formatted 3.5 diskette for each 8 sequences whose sequencer data files, including chromatograms, are requested. Reading chromatograms requires special software programs. Please submit the diskette with your samples, not when you retrieve the data. If you request data on a diskette, but do not provide a diskette, the Facility will provide you with a diskette at a charge of $1.^00.