I have just synthesized first strand cDNA samples from total RNA that I would like to use as templates in RACE PCR. The concentration of the cDNA is rather high though; I have four different samples of cDNA that range from 1,100 ng/uL to 1,800 ng/uL.
My protocol requires 2.5uL of template in the 50uL reaction.
1. What is the recommended concentration of cDNA I should use as my template?
2. How do I go about diluting my samples (calculation? water vs. buffer?) to obtain the desired concentration?