rom 60C to 95C.
DNA sequencing: We amplified fragments with PCR primers flanking the 300- to 600-bp BAC genomic sequence of interest in four family members, ran the PCR products in agarose gel electrophoresis, and purified the PCR fragments (QIAGEN Inc). Sequencing was performed with fluorescent dideoxy dye-primer on an ABI Prism 377 sequencer, and all results were visually inspected.
REFERENCES
- Cargill, C., et al. (1999) Nature Genetics 22: 231-238.
- Livak, K.J., et al. (1995) PCR Methods Appl. 4: 357-362.
- Chee, M., et al. (1996) Science 274: 610-614.
- Verge, C.F., et al. (1998) J. Clin. Invest. 102: 1569-1575.
- Tyagi, S., et al. (1998) Nature Biotechnol. 16: 49-53.
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