Patent ID: 8321140
Filing Date: 2012-11-27
Classification: C12Q

Abstract:
1. A method of determining an initial concentration (Q 0 ) of a target nucleic acid template in a sample, the method comprising the steps of: a) amplifying said target nucleic acid template and a plurality of standard templates, each of said standard templates present at a different, known copy number, in a nucleic acid amplification reaction regimen comprising a plurality of cycles of primer annealing, primer elongation and strand dissociation, wherein said reaction is performed using a reaction mixture comprising said target nucleic acid template and said plurality of standard templates, and wherein the efficiency of amplification, E, is similar for said plurality of standards and said target template; b) measuring, at plural cycles of said amplifying regimen, fluorescent signals from said target and each of said plurality of standard templates after nucleic acid species in an aliquot taken from the reaction mixture are separated by capillary electrophoresis, wherein said measuring generates a set of measurements for each said template; c) estimating a cycle threshold (C d) generating a standard curve by plotting C e) calculating an initial concentration (Q