Close
  Indian J Med Microbiol
 

Figure 5: Validation of wild type and mutant EGFR plasmid DNA by ddPCR: The 1D plot of QuantLife software showing the results of EGFR T790M mutation assay by ddPCR using wild-type and mutant plasmids as DNA templates. Two probes, one labeled with FAM (for mutant) and another with HEX (wild type), were used. Lanes B02 to D02 contained EGFR WT, #11011 plasmid DNA with varying concentration (20,000 copies, 10,000 copies and 5,000 copies respectively), whereas lanes E02 to G02 contained pBabeEGFR T790M, #32070 plasmid DNA varying concentration (20,000 copies, 10,000 copies, and 5000 copies respectively). The blue dots indicate the presence of FAM-labeled droplets positive for mutant allele (E02 to G02, upper panel), whereas green dots indicate the HEX-labeled droplets positive for wild-type allele (B02 to D02, lower panel). The channel amplitudes are plotted in Y-axis separately for FAM and HEX, whereas the events (number of droplets) are plotted in X-axis. The magenta line indicates the baseline and the black dots indicate droplets without any DNA

Figure 5: Validation of wild type and mutant EGFR plasmid DNA by ddPCR: The 1D plot of QuantLife software showing the results of EGFR T790M mutation assay by ddPCR using wild-type and mutant plasmids as DNA templates. Two probes, one labeled with FAM (for mutant) and another with HEX (wild type), were used. Lanes B02 to D02 contained EGFR WT, #11011 plasmid DNA with varying concentration (20,000 copies, 10,000 copies and 5,000 copies respectively), whereas lanes E02 to G02 contained pBabeEGFR T790M, #32070 plasmid DNA varying concentration (20,000 copies, 10,000 copies, and 5000 copies respectively). The blue dots indicate the presence of FAM-labeled droplets positive for mutant allele (E02 to G02, upper panel), whereas green dots indicate the HEX-labeled droplets positive for wild-type allele (B02 to D02, lower panel). The channel amplitudes are plotted in Y-axis separately for FAM and HEX, whereas the events (number of droplets) are plotted in X-axis. The magenta line indicates the baseline and the black dots indicate droplets without any DNA