This Facility provides a variety of molecular biology techniques, genotoxicity tests and general tissue and bacterial culture techniques, as well as training in these, to AHFCC members. The Facility is both responsive and proactive to the community it serves. Room 201 in Unit 2 is used for tissue culture and molecular biology and room 324 for bacterial work. The Facility has equipment needed for basic molecular biology: small centrifuges, speed vacs, electrophoresis equipment for general analysis sequencing, and blotting. A tissue culture hood and incubator are in the large of the two laboratories. Artek counters are used to score the Ames and UDS assays. To tube-based thermocyclers are available for PCR experiments. Other major equipment includes an oligonucleotide synthesis unit and Pharmacia SMART HPLC system for micro-purification of proteins. The senior investigators requested the following new services: expression of human and mouse genes using Clontech's 588 Atlas arrays; 32/P-post- labeling the DNA adducts and the single cell gel electrophoresis (COMET) assay for their repair, and Telomerase assay. The requested deep freezer will enable us to offer controlled long-term storage of plasmids, bacterial and mammalian cells. We shall work with the Histopathology Facility to develop analytical routines for image analysis using the requested fluorescence/transmission microscope upgrade and software system. Services currently available include the requested fluorescence/transmission microscope upgrade and software system. Services currently available include: The Ames Salmonella mutagenicity assays, with or without metabolic activation. The 7000 strains, containing defined mutations, will be made available. The hepatocyte DNA repair assays (Williams' test) uses human or rodent hepatocytes in vitro and rats in the in vivo/in vitro version. Oligonucleotide syntheses include carcinogen- modified nucleotides. DNA analysis via Souther blot hybridization and sequencing of enriched PCR analysis. Individual RNA analysis via Norther blot hybridization will be supplemented by the Clontech array. Protein analyses include Western blotting and protein-DNA interactions via gel retardation assays and micro-purification. We supply S9 and bacterial and mammalian cultures. The two excellent technicians provide an experienced and stable resource of experimental techniques and train other AHFCC personnel in tissue culture methods and molecular biology techniques.
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