ABSTRACT
The analysis of gene expression has become an important tool for understanding physiological and pathological processes, including development, plasticity, response to injury, carcinogenesis, regeneration, and death. Three approaches commonly used to systematically investigate changes in gene expression are plus–minus hybridization, subtractive hybridization, and reverse transcription messenger ribonucleic acids (mRNA) differential display. Reverse transcription mRNA differential display (mRNA-DD) is a polymerase chain reaction-based technique that allows a near-exhaustive comparison of gene expression from small samples of tissue obtained from multiple experimental groups. The essence of mRNA-DD is to use an anchored oligo-dT primer which anneals to the beginning of a subpopulation of the poly(A) tails of mRNAs for reverse transcription. The chapter utilises a modified version of mRNA-DD to look at potential differences in gene expression between sleep and waking. Primers are synthesized on a DNA synthesizer and purified by using a cleavage and deprotection kit according to the manufacturer’s instructions.
