The process typically involves the following key steps:
1. Template Preparation: A DNA template containing the gene of interest is cloned into a plasmid vector. This vector includes a promoter sequence recognized by RNA polymerases. 2. Transcription Reaction: The DNA template is mixed with a reaction buffer, RNA polymerase, and nucleotides (ATP, CTP, GTP, and UTP). The reaction is incubated at an optimal temperature to allow RNA synthesis. 3. RNA Purification: The synthesized RNA is purified to remove DNA templates, enzymes, and other contaminants. This can be achieved through methods like phenol-chloroform extraction and ethanol precipitation.
