The process of northern blotting involves several key steps:
Extraction of RNA: RNA is isolated from tissue samples using various extraction methods. Gel Electrophoresis: The extracted RNA is separated based on size through agarose gel electrophoresis. Transfer: The separated RNA is transferred from the gel to a membrane, typically made of nitrocellulose or nylon. Hybridization: A labeled probe (complementary to the RNA sequence of interest) is hybridized with the RNA on the membrane. Detection: The hybridized RNA is detected using various methods (e.g., autoradiography, chemiluminescence).
