The basis of PCR amplification or RNase security assay is, DNA and RNA probe will selectively hybridize with the molecules of nucleic acid of complementary sequence in a given sample which is commonly unknown. The most essential between Southern and Northern blotting is, southern blot is used after we have to uncover out the presence of a particular DNA fragment in a given sample. On the alternative northern blotting is a approach used for detecting RNAs which can be separated by the tactic of electrophoresis.
Southern blotting was launched by E.M. Southern in 70’s. In southern blotting what happens is, a DNA is extracted from the human cell which is of curiosity and then it is decrease into fragments with the help of restriction enzyme. Once these fragments are separated using the tactic of electrophoresis, they’re reworked into bands which comprise many copies of that DNA. We can also make these bands invisible via using explicit staining. Then these bands are transferred to nitrocellulose filter. Here the reply is made to cross by means of gel and filter to paper towel. This full course of produce nitrocellulose filter with DNA fragments transferred to the filter. This filter is then uncovered to radioactively labeled probes and this probe hybridize with a quick sequence of DNA. At ultimate filter is uncovered to X-ray film and the gene is acknowledged by a band on developed film.
Northern blotting was introduce by Alwine and his colleagues in 70’s. RNA is extracted on this course of from a given sample. Electrophoresis occurs and RNA is lastly separated by the size. After this, this RNA is transferred to the membrane and mounted to it with the help of UV gentle and heat. labeled probes are utilized on it which helps in visualization of labeled RNA over X-ray film. In Northern blotting, extraction of RNA is adopted by electrophoresis and then hybridization to a labeled DNA probe.
- Southern launched this technique of southern blotting. Northern blotting was launched by Alwine.
- Separation of DNA occurs in southern blotting, separation of RNA occurs in northern blotting.
- Denaturation should not be wished in northern blotting nonetheless it is required in southern blotting.
- Nitrocellulose filter membrane is utilized in southern blotting nonetheless not in northern.
- DNA-DNA hybridization occurs in southern blotting. RNA-DNA hybridization occurs in northern blotting.
- Amino benzoyloxymethyl filter paper is utilized in northern blotting nonetheless not in southern.