Is it enough to use only three biological replicates for correct statistics PCR-array analysis or three biological replicates and two technical replicates or one biological replicate and three tech replicates?
Our study divided into a total of 3 main groups and 15 subgroups. Each main groups contains 5 subgroups. These subgroups are associated with time (1h, 2h,3h, 4h and 5h). Like this:
Group A: Control Group C: Treatment Group
subgroup I, (1h)...........3 rats subgroup I, (1h)...........3 rats
subgroup 2 (2h)...........3 rats subgroup 2 (2h)...........3 rats
..... .....
subgroup 5 (5h)...........3 rats subgroup 5 (5h)...........3 rats
Group B: Negative Control
subgroup I, (1h)...........3 rats
subgroup 2 (2h)...........3 rats
.......
subgroup 5 (5h)...........3 rats
My questions
3 rats in each group are biological replicates. (I think). Are biological replicates enough? Or are technical replicates necessary? How many is enough for technical replicates?
and
At the end of the project, I will have results of each animals' 88 gen expression. Located timing in each sub-group is important, too. What program should I use to compare differences in gene expression? And how can create a heat map?