DNASU Sanger Sequencing • 480.965.6795 | Email



Sequencing Sample Preparation for Submission Step-by-Step

Please follow this four step process to prepare and submit your samples to the sequencing facility

Step One: Sample Preparation
We ask for 10ul each of DNA Template and Primer. Template should be prepared as high-quality plasmid DNA or purified PCR product. We can also provide DNA mini-preps if submitting colonies or glycerol stocks. Contact us for more information. Template and Primer concentrations are listed here. The following primers are available from the facility free of charge:

Primer Name Primer Sequence
M13 Forward 5'-GTAAAACGACGGCCAGT-3'
M13 Reverse 5'-CAGGAAACAGCTATGACC-3'
T7  5'-AATACGACTCACTATAG-3'
T7 Terminator  5'-TATGCTAGTTATTGCTCAG-3'
SP6  5'-CATACGATTTAGGTGACACTATAG-3'
T3    5'-AAATTAACCCTCACTAAAGG-3'

Step Two: Array Your Samples and Primers
Template and Primers should be arrayed separately on a non-skirted 96-well plate (VWR Cat# 82006-636) and sealed with aluminum adhesive seals. We can provide you with plates, tubes and seals if needed. Please array your samples as shown in the diagram. Your primer plate should also match the appropriate wells.

Also clearly label your tubes or plate to indicate where sample 1 is located and place the order ID Number on the tubes as well. Please number your samples as shown in white (top to bottom, left to right, one to ninety-six):

image of 96 well plate

Step Three: Submit Your Online Order
Once your samples have been arrayed you can login and submit your order into the online LIMS system and into the sequencing queue.

Once logged-in select "Enter Individual DNA Sequencing Requests" or "Upload and Import File." From here you can either enter individual samples or upload a previously edited .txt file. The template for the upload file can be downloaded from this page as well.

Now just enter the number of reactions you want to sequence, select the service type you want and click "submit" and you'll go to the online "DNA Sequencing Request Form".

Enter your PO/Billing Number in the top field then enter your DNA Template Name, select DNA Type and Primer from the drop-down menus, and click "Submit" and your order will be entered into the queue. There is a 22-character limit for sample names. You may also enter any comments about the samples such as GC-Rich or Low DNA Concentration that you would like us to know about.

Step Four: Submit Your Samples to the facility
Samples can be dropped-off at the Biodesign Institute
Please label as:

CPD Steel/Sequencing
Biodesign Institute
Arizona State University
1001 S. McAllister Ave.
Tempe, AZ 85287-6401
480-965-6795

Samples can also be shipped to the above address. We hope to have campus-wide pick-up locations in the future for your convenience.

You will receive an email when your samples are complete and ready to download. Just follow the link in the email, login to dnaLIMS and download your results. Click here for more information about programs that allow you to view your results.