I want to know expression level of a certain genes I do have primers . I have few questions:
1. is it necessary for me to quantify RNA? if it is only 1 sample?
2. Is it necessary that i should use random hexamers and oligo dT primers for cDNA synthesis?
3. If i could use gene specific Primers for cDNA Synthesis , should I use both primer [forward and reverse]?
actually i am thinking that I could use RNA without quantification. {yes quantification also tells about purity and maximum amount of RNA that i could use during cDNA synthesis} but if I am using same RNA and same amount that should not give me false results of expression . i am also assuming that i could use gene specific Primers forward and reverse both in same reaction.
dear researchers what are your views please explain.
thank you for reading .