I did qRT PCR with cDNA as the templates. I did two times, just for my curious, first for producting cDNA I used oligo d(T) and the other with random hexamer.
The problem is the qRT PCR data of both experiments are very different.
For example, by using cDNA with oligo d(T), the expression of the gene is shown high expressed (up regulated). But by cDNA with random hexamer, the gene is down regulated. I did just like normal qRT PCR (three replicates and three biological replications). So i'm really confuse now.
Is there anyohe who can explain it or perhaps has the same experiences??
When people should choice hexamer or oligo d(T) as the primer for cDNA production?
thanks for your answer