ABSTRACT
This chapter details how to utilize polymerase chain reaction (PCR) to go from answering the fundamental question, “Does a particular treatment affect the expression of a gene of interest,” to the examination of transcriptional activation, tissue distribution, and reporter gene analysis. The examination of a particular treatment on gene expression is of fundamental interest to the molecular toxicologist. The concentration of the target and reference primers must be determined empirically. In situ Reverse transcription PCR gives the researcher the ability to examine gene expression on a cell-by-cell basis. A type of control molecule that can be used to minimize tube-to-tube variability in amplification efficiency. Normally an Internal Standard is a synthetic molecule that contains the same primer recognition sequences as the gene of interest. Most often an External Standard is a housekeeping gene that is used in a coamplification type of quantitation.
