Center for Protein and Nucleic Acid Research

PLEASE SUBMIT TEMPLATE AND PRIMER IN TWO SEPARATE TUBES.

Templates and primers should be submitted in separate 1.5 mL tubes (can be requested from our web site).

For each template reaction, please submit 25 pmol primer (one only) in a total volume of 5 uL in a separate tube from the template. (concentration is 5 uM, or approx. 30 ng/uL for a 20mer primer). If submitting a primer for more than one template, please make sure you submit 5 uL per template sample submitted. Label primers with your initials and a letter to designate forward or reverse sequencing (e.g., ptoF or ptoR or for more than two primers, ptoF1, ptoF2...ptoR1, ptoR2...etc).

Primers should have a Tm ~58C-62C, be targeted to non-GC rich sites and contain no self-structure (please use mFold in useful links to check for self- structure).

Please be sure to indicate clearly on the template tubes and on the order form which primer goes with which templates using the labeling system described as follows: The primer tube above is labeled ptoF or ptoR, so just add an "F" or an "R" to back end of your template sample name (e.g., pto125F and pto125R, or if you have more than two primers, pto125F1, F2...R1, R2...etc). Please avoid the use overly complicated sample names.
(dissolve primer in dH2O only, TE buffer will inhibit the cycle seq reaction).

Please submit template in a total volume of 10 uL. Template quantity will vary depending on the size of plasmid or PCR fragment (see the table below). Accurate quantitation of template is essential to a successful sequencing reaction!

(Elute template from spin-column in dH2O only, TE or other elution buffers can inhibit the cycle seq reaction).

We offer a selection of primers, including M13 forward and reverse. We will add them to your sample for you. See our list of primers.

Submit the on-line order form. If you do not receive a confirmation email, your order did not go through, please resubmit your order.

Please write the sample name on the side of the tube.

If there are any problems with your data, please read the comments we place in the PDF file at the top. If you have further questions concerning your data, e-mail: dnaseq@scripps.edu.