Components of a Transcription Reaction

To carry out a transcription reaction in a test tube, several components need to be added in the right composition and in the right order. As in many in vitro experiments, a suitable buffer solution has to be supplied, taking care of maintaining the correct pH, as well as supplying essential reagents such as magnesium, polyvinyl alcohol, and KCl. Next added is the DNA fragment that contains the promoter. The RNA Polymerase II and the general transcription factors are added next. These components are mixed and incubated to allow sufficient time for the formation of the pre-initiation complex at the promoter site. The transcription reaction begins by the addition of a solution containing the ribonucleotides necessary for RNA synthesis. The final volume of a typical transcription reaction in Dr. Dvir’s lab is 35 microliters (very small indeed).

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