WM3211 Purified Genomic DNA

Genomic DNA was purified from cells using genomic DNA preparaton kit according to manufacturers instruction. DNA was diluted to 200 ng/µL in TE buffer (0.01 M Tris Chloride, 0.001 M EDTA, pH 7.6). Concentration was determined at A260 using nanodrop ND-1000. DNA was prepared from cell line WM3211. WM3211 is a tumorigenic (VGP) primary melanoma cell line with competence for metastasis. These cells display fibroblastic morphology in culture. This cell line was established from a metastatic site (lymph node) in a 34-year-old female. This cell line contains a L576P mutation at position 576 in the c-KIT gene. The L576P mutation results in an amino acid substitution at position 576 in KIT, from a Leucine (L) to a proline (P). This mutation occurs within the juxtamembrane domain. Mutant KIT proteins have increased kinase activity and transforming activity in vitro. This cell line is wild type for BRAF, PTEN, N-RAS, and CDK4. WM3211 cells produce xenograft tumors when injected into immunocompromised mice.