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New Protocols for Isolating High- Molecular-Weight Genomic DNA

od cells by centrifugation, then wash the pellet with NaCl (0.85%) solution.7 The pellet may be stored at 80C. When ready for use, thaw the pellet, and isolate and purify the DNA following the standard kit protocol.

Bone Marrow: Inject lysis buffer through two mouse femurs or one rat femur, and incubate the lysate on ice for 10 minutes; centrifuge and digest/dialyze per the standard kit protocol.

Cultured Cells: Suspend cells (1 to 5 x 106) in lysis buffer, eliminate the coarse disruption step, and follow the standard procedure.

REFERENCES

  1. Monforte, J.A., et al. (1995) Environ. Mol. Mutagen. 23 (Suppl. 23): 46.

  2. Rogers B.J., et al. (1996) Environ. Mol. Mutagen. 27 (Suppl. 27): 57.

  3. Short, J.M., et al. (1988) Fed. Proc. 8515a.

  4. Kohler, S.W., et al., (1991) Proc. Natl. Acad. Sci. USA 88: 7958-7962.

  5. Jakubczak, J.L., et al., (1996) Proc. Natl. Acad. Sci. USA 93: 9073-9078.

  6. Rogers, B.J., et al. (1996) Strategies 9 3: 73-74.

  7. Sambrook, J., et al. (1987) Molecular Cloning: A laboratory Manual,Second edition. Cold Spring Harbor Press. New York. p. 9.17.

  8. Rogers, B.J., (1998) Environ. Mol. Mutagen. 31 (Suppl. 29): 12.

* U.S. Patent Nos. 5,347,075 and 5,824,287 and 5,589,155; European Patent No. 0289121;Japanese Patent No. 2618973
** U.S. Patent No. 5,188,957


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