How much RNA should I use in my cDNA reaction?
We recommend using between 1-2 ug of RNA in your cDNA reaction. You MUST start with the same amount of total RNA in each sample. A first strand cDNA sysnthesis reaction is sufficient. You can buy cDNA reaction kits from Invitrogen, Applied Biosystems, Qiagen, Bio-Rad, amongst many others. Oligo d(T), Random hexamers, or a combination of the two can be used to prime the reverse transcription reaction EXCEPT if you choose to use 18s rRNA as an endogenous control. 18s rRNA requires the use of random hexamers since it lacks a poly A tail. To ensure all genomic DNA has been removed, a (-)RT reaction should be run.
