WM852 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 WM852. WM852 is a metastaic melanoma cell line with epithelial morphology. This cell line was established from the metastatic site (abdomen) in a 63-year -old male. This cell line contains a Q61R mutation at position 61 in the N-RAS gene. The Q61R is the most common NRAS mutation found in melanoma that is thought to occur due to UV and radiation exposure. This mutation leads to production of a constitutively active N-RAS protein that directs cells to grow and divide constantly. This cell line also expresses PTEN loss of function including hemizygous deletion and is wild type for BRAF, c-KIT and CDK4. WM852 cells produce xenograft tumors when injected into immunocompromised mice.