Gaussian decomposition of high-resolution melt curve derivatives for measuring genome-editing efficiency.

ABSTRACT: We describe a method for measuring genome editing efficiency from in silico analysis of high-resolution melt curve data. The melt curve data derived from amplicons of genome-edited or unmodified target sites were processed to remove the background fluorescent signal emanating from free fluorophore and then corrected for temperature-dependent quenching of fluorescence of double-stranded DNA-bound fluorophore. Corrected data were normalized and numerically differentiated to obtain the first derivatives of the melt curves. These were then mathematically modeled as a sum or superposition of minimal number of Gaussian components. Using Gaussian parameters determined by modeling of melt curve derivatives of unedited samples, we were able to model melt curve derivatives from genetically altered t

SUBMITTER: Zaboikin M 

PROVIDER: S-EPMC5754072 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature