WM46 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 WM46. WM46 is a metastatic human melanoma cell line that displays a pigmented fibroblastic morphology. This cell line was established from left chest & abdomen in a 43-year-old female patient with level IV superficial spreading melanoma and a tumor thickness of 1.44. This cell line features the specific V600E (Val600Glu) mutation at codon 600 in the BRAF gene. This mutation causes constitutively active kinase activity and activation of MEK and ERK signaling pathway. This cell line expresses PTEN loss of function including hemizygous deletion, and R24C mutation at position 24 in the CDK4 gene. The R24C mutation results in an amino acid substitution at codon 24 in CDK4, from an arginine (R) to cysteine (C). This mutation abolishes the ability of CDK4 to bind to p16(INK4a). WM46 cells produce xenograft tumors when injected into immunocompromised mice.