I do have 3 conditions, Basal, drug A 1uM 6 hours, drug A 1uM for 24 hours.
I would like to confirm the results of gene expression after basal and treatments with a specific drug. So first, I have to do RNA extraction, cDNA synthesis and qPCR. In other words, I want to make it clear there is, or there is no stimulation of the tested
gene because I got fold change between 1.2 and 2, and I know this is not enough in order to Conclude the effect of treatment
on X gene. So I decided to do quadruplicate of each condition- that means I will end up with
12 samples, I am confused is it better to plate in 6 well plate or 6 cm dish In order to have enough RNA? And after extraction of DNA
I will end up with 12 samples.,
I want to test 4 genes. So in qPCR, I will do it in duplicate then I will end up with 12×2=24 Samples.
12 samples need RNA extraction and cDNA synthesis ( table)
Then to run qPCR in (96 wells)
My questions are
1- IS this protocol will solve my drought regards the stimulation of genes (fold change)?
2- Is it better to plate in 6 well plate or 6 cm dish In order to have enough RNA?
3- 2- do the biological quadruplicate are correct in this way?
4- Do I distribute the technical duplicate samples and gene (yellow) correctly in 96 well plate for qPCR?
Thanks in advance.
