Director: Paul R. Kinchington, PhD
Molecular Biology is an essential component of virtually all biological research, particularly in light of the availability of a large volume of sequence information and the ready ability to manipulate it for examination of protein function. Molecular Biology is also a critical component for the follow up to modern genomic and proteomic applications. There has been an explosion in the variety of molecular applications and kits for performing them, and most are constantly changing and developing. As such, molecular biology applies to high throughput assays, manipulation and control of gene expression, mutational analysis of viruses, the host cell or the host animal. It is also a crucial component of most gene knockout and suppression strategies.
The Molecular Biology Module is housed in two rooms (350 sq. ft., and 100 sq. ft.) on the 10th floor of the Eye and Ear Institute.
The space includes a separate dark room for core equipment, phosphor imaging and UV gel analysis. Additional equipment required used by the module (ultra and high speed centrifuges, beta counter, etc) are located in a core instrument room, which is located approximately 30 feet from the Molecular Biology Module on the 10th floor. In general, work requests are prioritized on a first come-first served basis for participating NEI-funded investigators; but consideration is also given to small projects requiring immediate attention, investigators seeking pilot data for new vision-related grants, and vision-related research projects that are funded through other NIH institutes.
Major equipment includes hybridization ovens, several vertical and slab electrophoresis apparatuses and the power packs to run them, gel drying apparatus, Stratagene DNA cross-linker used in hybridization studies, UV transilluminator, and digital camera system, cooling and two shaking water baths, refrigerated micro centrifuge, an autoclave, high speed and ultracentrifuges, spectrophotometer and analytical balances. BioRad GS625 Phosphorimager, BioRad EthBr/fluoresce Max Gel and chemiluminescent documentation system, BioRad I cyclker PCR machine, PC computer (networked), and Gene pulser for bacterial transfections. In addition, the module has had access to the electrophoresis equipment of Dr. Kinchington, and has access to P2 tissue culture facilities, high speed and ultra-centrifuges, Speedvac apparatus, -20 and -70 freezers and cold storage, and scintillation counters, all of which are housed in common facilities of the Ophthalmology Research Center and are available for common use on an equally-shared basis.
- Preparation and analysis of DNA and RNA
- Preparation and screening analysis of phage libraries and generation of subtractive libraries
- DNA mutagenesis and expression
- DNA:protein, and protein:protein interactions
- RNA mapping and accurate quantification
- Virus vector design and derivation
Paul Kinchington, PhD
EEINS-1014, 203 Lothrop Street
Pittsburgh, PA 15213