WM3670 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 WM3670. WM3670 is a metastatic human melanoma cell line with epithelial morphology. This cell line features the G469E mutation at codon 469 in the BRAF gene. The G469E mutation results in an amino acid substitution at position 469 in BRAF, from a glycine (G) to a glutamic acid (E). This mutation occurs within the highly conserved GXGXXG motif of the kinase domain. Most mutant BRAF proteins have increased kinase activity and are transforming in vitro. This cell line contains a G12D mutation at position 12 in the N-RAS gene. The G12D mutation results in an amino acid substitution at position 12 in N-RAS, from a glycine (G) to an aspartic acid (D).