Options PageNo=1 NoDate;
Title 'Real-time PCR Based Transgene Copy Number Determination';

Data T2;
 Infile 'X:\' ExpandTabs; /*Please input your data directory here*/
 Input Cat $ Gene $ Con Ct;
 ;
Run;

/*Logarithm 2 based data transformation.*/
Data T3;
 Set T2;
 LX=Log(Con)/log(2);
Run;

Proc Print Data=T3; Run;

/*Simple linear regression for amplification efficiency estimation*/
Proc Reg Data=T3;
 By Gene;
 Model LX=Ct;
 Test Ct=-1;
Run;

/*ANOVA model*/
PROC MIXED data=T3 order=Data;
               Class Cat;
               Model Ct = LX|Cat /SOLUTION cl;
               Contrast  'DeltaDeltaCt' Cat 1 -1;
               Estimate 'DeltaDeltaCt'  Cat 1 -1/cl;
Run;