Performance of Mutation Assays using RNA or DNA Extracted from Cell Lines.

aThe CT and standard deviation (SD) comparing a 50/50 wild type/mutant cell line mixture (32 ng each) with wild type alone (32 ng) using the indicated assay. Wild type cell lines used were either COLO 320 or HeLa, depending on availability at the time. Cell line SW480 is homozygous for Mut1 (G216T); all other cell lines are heterozygous for the indicated mutations. Linearity of all assays ranged from 0.992–0.999. Efficiency for all assays ranged from 92%–116%. ΔCT is the difference between the CT obtained from Wild Type Cell Line CT – Mixed Cell Line CT. Selectivity was measured as described in Results.
bThe CT and standard deviation (SD) comparing a 50/50 wild type cell line DNA/mutant synthetic DNA (15,625 copies each) mixture with wild type DNA alone (15,625 copies) using the Mut6.1 assay. Synthetic mutant DNA was used for Assay Mut6.1 as no cell line carrying this mutation was available. Limiting Dilution Assay analysis was used to determine the number of copies of synthetic template as well as the number of wild type Kras alleles in HeLa DNA.