Tissue Homogenization and DNA Extraction

1. Tissue.

2. Liquid nitrogen.

3. Mortar and pestle (Fisher Scientific, Houston, TX).

4. Proteinase K (Gibco-BRL, Gaithersburg, MD).

Fig. 3. Mutation of IGFIIR. H54 (N and T), normal and tumor DNAs from a patient with ulcerative colitis-associated colorectal cancer; 4854, G28, JG613, JG831, and IG15 (N and T), DNAs from patients with gastric cancer; AC31and AC44 (N and T), DNAs from patients with sporadic colorectal adenocarcinoma; AC41 (N and T), DNAs from a patient with HNPCC. Mutations were demonstrated using primer set R4, except for 4854 N,T (with a mutation demonstrated using primer set R5). An abnormally migrating band, located just above or below the wild-type band, is visible in each of the tumor (T) lanes.

Fig. 3. Mutation of IGFIIR. H54 (N and T), normal and tumor DNAs from a patient with ulcerative colitis-associated colorectal cancer; 4854, G28, JG613, JG831, and IG15 (N and T), DNAs from patients with gastric cancer; AC31and AC44 (N and T), DNAs from patients with sporadic colorectal adenocarcinoma; AC41 (N and T), DNAs from a patient with HNPCC. Mutations were demonstrated using primer set R4, except for 4854 N,T (with a mutation demonstrated using primer set R5). An abnormally migrating band, located just above or below the wild-type band, is visible in each of the tumor (T) lanes.

5. TE buffer: Tris-HCl base 10 mM and EDTA 1 mM, adjusted to pH 8.0 (Sigma, St. Louis, MO).

7. Saturated phenol buffer, pH 8.0 (Gibco-BRL).

8. Chloroform (Fisher Scientific).

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