Introduction to DNA sequence

The Molecular Core is currently doing all its sequencing through Agencourt. This gives our users access to the latest sequencing technology and competative pricing.

This service provides quick turnaround sequencing of purified individual samples, including PCR amplicons, low copy plasmids and high copy plasmids. Single reads are performed on each sample using customer-provided primers or Agencourt-provided universal primers. Samples can be submitted in 2D bar coded tubes or 96-well plates. Primers can be mixed in with the template or sent in separate plate(s) or tube(s). Data is posted to our secure FTP site 24 hours after sample receipt, or 48 hours if the samples need to be reworked. Note that an increased turn around time will result if Agencourt is required to purify the samples or design primers.

Agencourt will design and synthesize primers to obtain full length sequencing of PCR amplicons, plasmid inserts or entire plasmids. You can select single strand or publication quality (double strand) coverage. Turn around time is approximately 5-7 business days per round of walks, depending on the project, but may be expedited by sending reference sequence early. Agencourt will deliver a contiguous consensus sequence in FASTA format. You can submit 100 μL glycerol stocks in either 2D tubes or 96-well plates.

Automated sequencing is less tolerant to variations in sample and primer impurities than manual sequencing. Primer design has to be optimized for sequencing. Just because a primer works to amplify a PCR band does not necessarily guarantee that it will be a good DNA sequencing primer. See sequencing trouble shooting below.

In Tubes: For QuickLane, please send DNA in a 2D bar-coded tube. For Primer Walking or unpurified plasmid template QuickLane Samples, please submit 100 μl of glycerol stock.

In Plates: Write plate name on the plate and Agencourt will barcode the plate when it is received at our facility and associate barcode with naming on plate.

Primers: Custom primers may also be submitted in tubes or 96-well plates. For pre-designed primers, please submit primer sequence. If you want Agencourt to design and synthesize primers for you, please submit reference sequence in FASTA format. Turn around time can be expedited if you send sequence 3 business days prior to template shipment.

You can always check the most current version of our Sample Submission Guidelines, which are posted in the Agencourt DNA Sequencing section of the UCHC Molecular Core website.

Submitting Template and Primer Mixed: The table below indicates concentration and volumes of DNA template and primer needed when mixed together. Please consult the Sample Submission Guidelines for complete information.

Submitting Template and Primer Unmixed: The table below indicates concentration and volumes of DNA template and primer needed when they are not mixed together. Please consult the Sample Submission Guidelines for complete information.

Submitting Unpurified Plasmid Glycerol Stock: Agencourt offers purification of plasmid glycerol stocks. Submit 100µL of plasmid glycerol stock per sample in 2D tubes or 96-well plates. (To submit in plate format, the customer must submit 8 samples or more). Please consult the Sample Submission Guidelines for complete information.

Agencourt Supplied Primers: The table below contains a list of all Agencourt-supplied primers. Please call your Account Manager if you require additional frequently-used primers to be added to this list:

Setting Up a New Account:

Go to the Molecular Core Website, which can be found at the following URL in your web browser*: http://mc.uchc.edu/mc/index_molecore.htm
Log into the UCHC Molecular Core Website, using your current username and password.
Click on the “Agencourt DNA Sequence” link from the menu on the left.
In the Table labeled “The Steps to Use Agencourt DNA Sequencing Services”, subsection “The steps before using Agencourt DNA Sequencing Services”, click on Item #1, “New Agencourt Account”.
You will now enter the Agencourt Project Submission Form (PSF) Account Registration Page. Here you will enter your personal account information. Note: please use your @uchc.edu e-mail address. When your information is complete, click the “Submit” button.
You will return to the Molecular Core Website, and click #2, “Save Agencourt Account Information”. Here you will add your Agencourt username and password to your UCHC Molecular Core profile. Note, you will only have to do this step once.

New Project Creation:

Go to the Molecular Core Website, which can be found at the following URL in your web browser*: http://mc.uchc.edu/mc/index_molecore.htm
Log into the UCHC Molecular Core Website, using your current username and password.
Click on the “Agencourt DNA Sequence” link from the menu on the left.
In the Table labeled “The Steps to Use Agencourt DNA Sequencing Services”, under the subsection “The Steps to Use Agencourt DNA Sequencing Services”, click on # 2 “New Order From Molecular Core”.
Choose the appropriate “FRS Account Number”. Complete the form by entering the project details (project name, reaction number, etc.) and then click “Submit”.
Look for the “Agen” blue hyperlink (located to the left of the Project Name), and click on it to get to the Agencourt Online Project Submission Form (PSF).
Once inside the the PSF, complete your project information (i.e. template, primer, etc.).
In the P.O. field, enter “UCHC” (confidential).
Click the “Submit” button. Print the Summary Page, and enclose it with your samples.
For questions regarding the Agencourt PSF, please contact your Agencourt Team (see Page 3 below) or call Customer Service at 800-361-7780 or the UCHC Molecular Core at 860-679-3720.

* If you have a pop-up blocker, you must first disable it

Back to Top

DNA Sequencing
	Our Sequencing Schedule	Sample Preparation	Sample Submission	Use Of This Website for Sample Submission