The BioRad Opus is our quantitative PCR system. To use, first setup an account at BR.io. Once you have an account setup, link your account to our thermocycler. Instructions can be found at BR.io.
Our laboratory has an SOP for quantitative PCR. Please see the linked protocol for performing qP
There are several approaches to analyzing qRT-PCR results. Below are some methods used by the Nicholas Lab and resources for performing the analysis.
* Delta Delta CT method – The delta-delta Ct method, also known as the 2–∆∆Ct method, is a simple formula used in order to calculate the relative fold gene expression of samples when performing real-time polymerase chain reaction. These calculations can be performed in an excel spreadsheet or with the aid of the CFX connect software at the data analysis station in lab.
*geNorm method – geNorm is a popular algorithm to determine the most stable reference (housekeeping) genes from a set of tested candidate reference genes in a given sample panel. From this, a gene expression normalization factor can be calculated for each sample based on the geometric mean of a user-defined number of reference genes. The geNorm method can take into account multiple housekeeping genes.