NORTHERN BLOTTINGS Meaning and
Definition
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Northern blotting is a laboratory technique used in molecular biology to study and analyze specific RNA molecules present in a sample. Named after its counterpart technique, Southern blotting, which is used to detect specific DNA molecules, Northern blotting focuses on the detection and analysis of RNA.
The process of Northern blotting involves separating RNA molecules according to their size through gel electrophoresis. The RNA sample is loaded onto an agarose gel and an electric current is applied, causing the RNA molecules to migrate through the gel based on their size. Once separated, the RNA molecules are then transferred (or blotted) onto a solid membrane, typically made of nylon or nitrocellulose.
The next step in Northern blotting involves hybridization, where a labeled single-stranded DNA or RNA probe is added to the membrane. This probe is complementary to the specific RNA molecule being studied and will bind to it. The probe can be radioactively or chemically labeled, allowing for the detection and visualization of the RNA molecule on the membrane.
By using Northern blotting, scientists can determine the abundance of specific RNA molecules in a sample and study their expression patterns. This technique has been invaluable in various areas of molecular biology, including gene expression analysis, identification of RNA isoforms, and studying RNA post-transcriptional modifications. Although newer methods such as reverse transcription-polymerase chain reaction (RT-PCR) and RNA sequencing have gained popularity, Northern blotting remains a useful tool for studying RNA molecules in a targeted and quantitative manner.
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