What is the Difference Between Northern Southern and Western Blotting?
🆚 Go to Comparative Table 🆚The main difference between Northern, Southern, and Western blotting techniques lies in the type of molecule they are designed to identify:
- Northern Blotting: This technique is used to detect and identify specific RNA sequences in a sample of mixed RNAs. The process involves RNA isolation, electrophoresis, and transfer of RNA bands to a membrane. Northern blotting can be utilized in gene expression research and disease diagnosis.
- Southern Blotting: This method is used to detect a specific DNA sequence in a sample of mixed DNA. The protocol begins with DNA extraction, followed by electrophoresis, transfer of DNA bands to a membrane, and identification of specific sequences using DNA probes. Southern blotting is useful for studying genetic diseases and identifying specific DNA sequences.
- Western Blotting: This technique detects specific proteins in a sample. The process involves protein isolation, electrophoresis, and transfer of protein bands to a membrane. Western blotting is an immunoblotting technique, which means it uses antibodies to identify the target protein. It is often used to study protein expression, function, and interactions.
In summary, the primary difference between Northern, Southern, and Western blotting techniques is the molecule they target: RNA for Northern blotting, DNA for Southern blotting, and proteins for Western blotting. While all three techniques involve electrophoresis and transfer to a membrane, they use different gels and methods for detection.
Comparative Table: Northern Southern vs Western Blotting
Here is a table comparing the differences between Southern, Northern, and Western blotting techniques:
| Technique | Target Molecule | Detection Method | Electrophoresis | Application |
|---|---|---|---|---|
| Southern | DNA | Hybridization with radiolabeled or fluorescent probe | Agarose gel electrophoresis | Detect specific DNA sequences |
| Northern | RNA | Hybridization with cDNA or RNA probe | Formaldehyde agarose gel or glyoxal agarose gel electrophoresis | Detect specific RNA sequences |
| Western | Protein | Antibody-antigen binding | SDS-PAGE | Detect specific proteins |
Southern blotting is used to detect specific DNA sequences, Northern blotting for specific RNA sequences, and Western blotting for specific proteins. All three techniques involve electrophoresis, transfer to a membrane, and a detection step, but they differ in the type of molecule they target and the specific method used for detection.
- SDS Page vs Western Blot
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