Table 2. The analytic procedure of the example RT-qPCR data using SASqPCR.

Code#	SAS code for analysis of study example data*
1	Filename myresult “X:\qPCR\PCR_data.xls”; run;
2	proc import out = work.raw_ct datafile = “X:\qPCR\PCR_data.xls”dbms = Excel replace; sheet = “Raw_Ct”; run;
3	%Include “X:\qPCR\SASqPCR.sas”; run;
4	%Efficiency(10, 5);
5	%stability(0.95, “14-3-3e” “Act5C” “Appl” “CG13220” “Cyp1” “Ef1a48D” “Elav”“Exba” “Gapdh2” “GstD1” “Rap2l” “Robl” “RpL13A” “RpL32”“Sdha” “Su(Tpl)” “aTub84B” “eIF-1A” “l(3)02640”, 19);
6	%Optimization(“C05”, 19);
7	%Normalization(“C05”, “Exba” “Cyp1” “l(3)02640” “Appl” “14-3-3e”, 5);
8	%Exp_R(“Atg1” “Aß42” “CathD” “Hsp70” “InR” “Lamp1” “Rab5” “Tor” “Hu_tau”);

*The folder “X:\qPCR” in code #1, #2 and #3 needs to be changed to the appropriate path and filename so that SAS software can successfully access it. Input names of genes and samples must exactly match those in the original dataset. Please note that it is possible but not necessary to use the same Excel file to save the raw Ct data and exported results.