Rat Brain-Heart-Liver Control Mix Self-Self Hybridization

Eight “self-self” hybridizations were prepared as follows. Total RNA was isolated from each of three rat livers and brains, and equal mass amounts of each of these six samples were combined to create a mixed RNA population. Cy3- and Cy5-labeled cDNA was generated from this RNA mixture using an aminoallyl nucleoside analog indirect labeling protocol. Specifically, eight separate 20 microgram aliquots of the total RNA mixture were reverse transcribed into cDNA in the presence of aminoallyl-dUTP and the resulting cDNA was then coupled to either Cy3 or Cy5 mono NHS ester (four reactions with each label). Keywords: Self-Self Hybridization Two Color Experiment Dye-Swap Design with identical biological sample and 4 technical replicates.