<Department of Microbiology & Immunology, UT Health Science Center at San Antonio Double Helix rotating/>

Sequencing


Sequencing Project Form                        FAQs                        Primer Stock List

The NACF houses two ABI 3130xl instruments and sequencing and fragment analysis are offered on a service-for-fee basis with a range of services available to investigators at the HSC and surrounding scientific community.





Sequencing runs are done daily. All samples that are submitted by 11:00am will be processed the same day with results available the next working day in most cases. When workload exceeds our maximum capacity the turn-around times may vary between one and three days and users will be notified of the extended turn-around time.

Service

Description

Cost

Raw Data SEQ RXN

Cycle sequencing reaction (rxn) with one template, one primer, and ABI Big Dye Terminator chemistry is performed. Products are purified and analyzed by the 3130xl.

$14.00/RXN

Edited Data SEQ RXN

Cycle sequencing reaction (rxn) with one template, one primer, and ABI Big Dye chemistry is performed. Products are purified, analyzed by the 3130xl, and data is analyzed using Sequencer software. Data analysis may include: sequence editing, assembly of consensus sequences, and primer walking.

$16.00/RXN

HT SEQ RXNS
(48 or 96 rxns)

Rxn component (template + primer) aliquots are submitted in a core approved 96-well plate. Core performs cycle sequencing with ABI Big Dye chemistry. Rxn products are purified and analyzed by the 3130xl.  
*Price is dependent on volume and is negotiated on an individual basis and can be as low as $6/rxn.

$6.00 /RXN*

READY RXNS

User performs cycle sequencing rxns and submits products as purified lyophilized samples ready for analysis on the 3130xl.      *Price is dependent on volume and is negotiated on an individual basis and can be as low as $3/rxn.

$3.00/RXN*

Sample Quantification by OD

OD is read on the NanoDrop ND-1000 and a print-out with concentration and 260/280 ratio is provided.

$1.00/Sample

Sample Concentration Correction/Dilution

-

$1.00/Sample

Color Chromatogram Printout

-

$1.00/Page


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Quality Sequence Requires Quality Template
 The quality and quantity of your DNA are critical to achieving optimal results. Cycle sequencing is sensitive to impurities in the template as well as to the concentration of template in the reaction mixture. It is crucial that DNA samples are clean, free of alcohol and salts, and have an accurate concentration. Recommendations for preparation and care of samples can be found in the FAQs.

Template Type

Amount

Concentration

Plasmid

0.8 - 1 μg / rxn

0.1 μg / μl

PCR Product

80 - 100 ng / rxn

10 ng / μl

Primers

3.2 pmol / rxn

1 pmol / μl

 

Primer Considerations:

Tm should be between 52-56°C. Avoid a Tm < 50°C.
Try to design primers with at least 50% GC content.
Avoid strings of four or more of the same base.
Design primers at least 30 bases from the region of interest.

 

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On Campus:
  1. Complete a sequencing project form to submit with the samples to be sequenced.
  2. Legibly label sample and primer microcentrifuge tubes. Please limit names to no more than 10 alphanumeric characters and do NOT use symbols, dashes, or other non-alphanumeric characters.
  3. Number the side of your sample tube(s) with the number corresponding to the line number on the sequencing project form.
  4. Deliver samples/primers to the lab (4.059V) or to the Sequencing Drop Box located in the refrigerator across from our lab. Please enclose your project form along with samples in a ziplock bag which is provided in the drop box.
*The Sequencing Drop Box is accessible to users all the time and is checked frequently during normal operating hours.

Off Main Campus:

  1. Follow steps 1-3 of the above procedure.
  2. Enclose form and securely close samples/primers in a 9"X12" envelope labeled:
             Nucleic Acids Core Facility
             Dept of Microbiology Rm 4.059 V
  3. Send through campus mail.
* Keep in mind that campus mail is delivered only once a day at 3pm.

Non-UTHSCSA:

  1. Follow steps 1-3 of the above procedure.
  2. Sample/primers should be closed securely and placed in a small ziplock bag. Enclose samples and form in a 9"X12" envelope or a small disposable cooler. (Samples may be sent on ice if desired, but please ensure that the project form remains dry.)
    3. UTHSCSA
    Attention: Nucleic Acids Core Facility
    Dept of Microbiology Rm# 4.059 V
    7703 Floyd Curl Drive
    San Antonio, TX 78229-3900
* Please allow the appropriate time for selected shipping method.

*HT Sequencing, Ready Reaction, & Fragment Analysis
Please call or come by the lab for specific instructions for submission.


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