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 . 

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