Baylor Remains Open

Baylor College of Medicine and its clinics will be open Monday, Nov. 28. We are closely monitoring the City of Houston's boil water advisory and will provide updates as needed.


Adenovirus Production Outline

Steps Performed By Adenovirus Production Outline
  • Obtains pShuttle plasmid from the Gene Vector Core
  • Clones gene of interest into pShuttle
  • Transforms plasmid and selects for Kanamycin-resistant colonies
  • Screens mini-prep DNA from colonies by restriction digests
  • Sends at least 100ug of the plasmid to the Gene Vector Core
Gene Vector Core
  • Expression cassette excised from pShuttle
  • Expression cassette ligated into the Adeno Viral DNA
  • Ligation reaction digested with Swa I and transformed into Stbl2 cells
  • Ampicillin-resistant colonies screened by restriction digests
  • Maxi-prep of the final plasmid are made
  • Adenoviral DNA digested with Pac I
  • HEK 293 cells transfected and monitored for plaque formation
  • Plaques picked and small-scale expansion performed
  • Positive samples sent to investigators to test and select sample for final expansion & purification
Gene Vector Core
  • Medium-scale expansion of investigator's selected sample for production of lysate
  • Viral lysate used to infect cell factory
  • Large-scale expansion harvested and purified on two CsCl gradients
  • Smaple desalted, aliquoted and frozen
  • QA/QC testing
  • Virus shipped by the Gene Vector Core to the investigator