Analysis of qPCR data by converting exponentially related Ct values into linearly related X0 values.
A common method for calculating results from qPCR experiments is the comparative Ct method, also called the 2(-ΔΔCt) method. However, several assumptions are included in the 2(-ΔΔCt) method and standard statistical analyses are not directly applicable. Here, we describe a different method, the X(0) method, for result calculations and statistical analysis from qPCR experiments. The X(0) method differs from the 2(-ΔΔCt) method by introducing a conversion of the exponentially related Ct values into linearly related X(0) values, which represent the amount of starting material in a qPCR experiment. Results calculated by the X(0) method are illustrated for qPCR experiments with technical and biological replicates, including procedures to calculate standard deviations. Incorporation of primer efficiencies in calculations by the X(0) method is also described. Altogether, the X(0) method constitutes a very simple and accurate alternative to the 2(-ΔΔCt) method for result calculations from qPCR data.
Thomsen
Rune
R
Department of Human Genetics, The Bartholin Buildin, Aarhus University, DK-8000 Aarhus C, Denmark. Rune@humgen.au.dk
Sølvsten
Christina Ane Elisabeth
CA
Linnet
Toke Elbek
TE
Blechingberg
Jenny
J
Nielsen
Anders Lade
AL
D000818:Animals; D001921:Brain / Q000378:metabolism; D019295:Computational Biology; D003627:Data Interpretation, Statistical; D015870:Gene Expression; D016259:Genes, myc; D007668:Kidney / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D016133:Polymerase Chain Reaction / Q000706:statistics & numerical data*; D012333:RNA, Messenger / Q000235:genetics
D012333:RNA, Messenger