The University of Texas Medical School at Houston
Department of Microbiology and Molecular Genetics
The Department of Microbiology and Molecular Genetics

Facilities

The Department of Microbiology & Molecular Genetics at the University of Texas Medical School at Houston has many facilities.


DNA Sequencing Service

The MMG Core Facility is now located in room MSB1.004 and provides automated DNA sequencing services for all interested researchers within the UT Health Science Center as well as other academic institutions.

Electron Microscope

The MMG department houses a JEOL JEM-1400, a 120 keV cryo-transmission electron microscope with electron tomography capabilities. See the Electron Microscope Facility website for more information.

Biacore System

The MMG Core Facility houses a Biacore 2000 instrument, which employs state-of-the-art surface plasmon resonance technology and microfluidics for the real-time analysis of macromolecular interactions. Contact the Core Facility to obtain instrument training and to discuss use of the Biacore system (713-500-5455 or core@uth.tmc.edu)

Real Time PCR

The MMG Core Facility houses a ABI 7500 Real Time PCR System (96 well plate). Real time PCR is available at $25 per run. This price includes only the run. Reagents and consumables are not included in this price. Contact the Core Facility for more infromation (713-500-5455 or core@uth.tmc.edu)

Sequencing Sample Preparation Guidelines

  1. Prepare both the primer and the template DNA in deionized water.
  2. Follow link on Core homepage for a list of vector primers we can provide.
  3. Dilute your custom primer to a concentration of 5 pmol/µl (5 µM). At least 3 µl is required for each reaction. Please provide a little extra.
  4. 0.5 µg of ssDNA or 1.0 µg of dsDNA at a concentration of 200 ng/µl is required for each reaction (except as noted below).
  5. Please indicate on the submission form if the template to be sequenced is larger than 15 kb or smaller than 1kb.
  6. To sequence clones much larger than 15 kb (e.g., lambda or BAC clones), please increase the template concentration to 500 ng/µl and provide your custom primer at a concentration of 30 pmol/µl (30 µM).
  7. For PCR products and other templates that are shorter than 1.0 kb, the amount of template needed is determined by the length. For each reaction, 25 to 50 ng/100 bp is recommended in a final volume of 10 µl.
  8. There is no need to aliquot your template or primer if it is being submitted for multiple reactions.

Sequencing Primers Provided

Primer Sequence (5' to 3')
M13 Forward (-21) TGT AAA ACG ACG GCC AGT
M13 (-40) GTT TTC CCA GTC ACG AC
M13 Reverse GGA AAC AGC TAT GAC CAT G
T3 AAT TAA CCC TCA CTA AAG GG
pBST3 CCT CAC TAA AGG GAA CAA AAG C
T7 GTA ATA CGA CTC ACT ATA GGG C
EXT7-1 TAA TAC GAC TCA CTA TAG GGC GA
EXT7-2 TAA TAC GAC TCA CTA TAG GGA GA
EXT7-3 TAA TAC GAC TCA CTA TAG GGG AA
T7-Terminator GCT AGT TAT TGC TCA GCG G
SP6 CAT ACG ATT TAG GTG ACA CTA TAG
SK CGC TCT AGA ACT AGT GGA TC
KS TCG AGG TCG ACG GTA TC
CMV-F CGC AAA TGG GCG GTA GGC GTG
BGH-R TAG AAG GCA CAG TCG AGG
 

Obtaining Your Results

Currently we can provide the ABI file and a simple text file on your floppy, zip (100) disc, CD or (USB) flash drive, and a hard copy of the electropherogram.

To open your ABI file, download Finch TV (for Linux, PC, or Mac OSX, click here to install), Chromas (for PC, click here to install), or EditView (for Mac OS9, click here to install) depending on which is compatible with your computer. All three programs are free.

Technical Sequencing Information

    For DNA sequencing, we employ:
  • chemistry: cycle sequencing, dye terminator (Unidirectional PCR reaction using terminators labeled with different fluorescent dyes. Our current standard dye is Applied Biosystems BigDye® Terminator v.3.1)
  • Post-PCR cleanup: paramagnetic beads (Agencourt)
  • Separation: capillary electrophoresis (ABI 3130 XL, 80cm array)