Abstract

The tissue culture core component will provide three principal services for projects 1-4: (1) It will obtain, passage, and maintain frozen stocks of primary cultures of human foreskin fibroblasts and keratinocytes; (2) It will provide appropriate growth-phase cells, both from primary cultures and immortalized cell lines as necessary for all experiments; (3) It will order, inventory, and quality-control test all media, sera, and growth additives. Concomitant with provision of cells for experiments, the tissue culture core will optimize growth conditions, determine doubling times and lag/log growth periods, and perform cytotoxicity and growth- delay assays using the mutagens at concentrations and schedules indicated by the damage mapping experiments. Because of the commonality of cell types and experimental protocols of projects 1-4, performing tissue culture functions in a core component will improve overall quality and the comparability of the data between projects, and will result in economy of effort and significant cost savings as well.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Program Projects (P01)
Project #
1P01CA069449-01A1
Application #
5209528
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
1
Fiscal Year
1996
Total Cost
Indirect Cost

  Publications

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Cloutier, J F; Castonguay, A; O'Connor, T R et al. (2001) Alkylating agent and chromatin structure determine sequence context-dependent formation of alkylpurines. J Mol Biol 306:169-88
Chen, J Z; Smith, L; Pfeifer, G P et al. (2001) Fluorescence-based directed termination PCR: direct mutation characterization without sequencing. Nucleic Acids Res 29:E17
Miao, F; Bouziane, M; Dammann, R et al. (2000) 3-Methyladenine-DNA glycosylase (MPG protein) interacts with human RAD23 proteins. J Biol Chem 275:28433-8
Bouziane, M; Miao, F; Bates, S E et al. (2000) Promoter structure and cell cycle dependent expression of the human methylpurine-DNA glycosylase gene. Mutat Res 461:15-29
Pfeifer, G P (2000) p53 mutational spectra and the role of methylated CpG sequences. Mutat Res 450:155-66
Szabo, P E; Pfeifer, G P; Miao, F et al. (2000) Improved in vivo dimethyl sulfate footprinting using AlkA protein: DNA-protein interactions at the mouse H19 gene promoter in primary embryo fibroblasts. Anal Biochem 283:112-6
Chen, J Z; Qiu, J; Shen, B et al. (2000) Mutational spectrum analysis of RNase H(35) deficient Saccharomyces cerevisiae using fluorescence-based directed termination PCR. Nucleic Acids Res 28:3649-56

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