What is the recommended primer for reverse transcription: Oligo(dT), random hexamer or gene specific primer?
In principle the usage of all primers is possible. Higher specificity and yield of transcribed product can be achieved when using gene specific primers. Starting amounts for an optimization of the experiment on the AmpliGrid for a one µL should be: – gene specific primer: 0.6 pmol for singleplex; 0.2 pmol for 10-plex – Oligo(dT): 3.5 pmol – random hexamer: 1-1.5 pmol Example for calculating the amounts out of primer concentrations: 0.6 pmol are 0.1 µL of a 6 µM primer solution (each sense and antisense) in water.
