5/9/2023 0 Comments Immunoblotting vs western blotSouthern blotting is used to identify specific gene sequences and in DNA fingerprinting. Southern blotting uses DNA probes while Northern blotting uses cDNA probes and Western blotting uses primary and secondary antibodies. Blotting Methodīoth Southern and Northern Blotting involve capillary transfer while Western blotting involves an electric transfer. Southern blotting involves Agarose gel electrophoresis, and Northern blotting involves denaturing formaldehyde agarose gel while Western blotting involves SDS PAGE. Southern blotting detects specific DNA sequences, Northern blotting detects specific RNA sequences, while Western blotting detects specific proteins. Northern blotting was developed by Alwine and his colleagues in 1979, and Western blotting was developed by George Stark’s group at Stanford University in 1979. Southern blotting was developed by Edward M. Southern blotting refers to a procedure for identifying specific sequences of DNA, while Northern blotting refers to an adaptation of the Southern blot procedure used to detect specific sequences of RNA by hybridization with complementary DNA and Western blotting refers to a blotting procedure used to identify specific amino-acid sequences in proteins. Each technique has applications in biotechnology and medicine.ĭifference Between Southern Northern and Western Blotting Definition.Each technique requires denaturation and blocking with the excess, corresponding macromolecule.The three steps of the blotting techniques are electrophoresis, transfer, and detection.Southern blotting, Northern blotting, and Western blotting are three blotting techniques used to identify a particular type of macromolecule within a sample.Similarities Between Southern Northern and Western Blotting Moreover, western blotting is the definitive test for Hepatitis B, Creutzfeldt-Jacob disease, Lyme disease, and Herpes. It can identify the number of proteins in a sample, presence of bacteria and virus in serum, and the presence of HIV antibodies in the serum. ![]() Western blotting is also called protein blot or immunoblotting. High stringency defined by the high temperature and low salt concentration increases the specificity of the hybridization while low stringency defined by low temperature and high salt concentration decreases the specificity. Stringency, which depends on the temperature and the salt concentration of the hybridization buffer, affect hybridization. The hybridization probe is short, 100-500 bp in size, single-stranded DNA molecule. Transfer – During the transfer, a nitrocellulose membrane is placed in contact with the gel and DNA bands move onto the membrane through the capillary action.ĭetection of Specific Sequences – The target sequence on the membrane is hybridized with the specific, labelled oligonucleotide sequence called the hybridization probe. The basic steps of a Southern blot are as below.Įlectrophoresis – It separates the DNA sample into distinct bands based on the size by gel electrophoresis. Southern blotting is the technique used to detect a specific DNA sequence in a mixture of DNAs. – Comparison of Key Differences Key TermsĪntibody, DNA, Electrophoresis, Northern Blotting, RNA, Proteins, Southern Blotting, Western Blotting What is the Difference Between Southern Northern and Western Blotting What are the Similarities Between Southern Northern and Western Blottingĥ. During blotting, the macromolecules are transferred onto a membrane from the gel and made to bind with a specific nucleic acid or antibody that aid in the detection. Southern, Northern, and Western are three blotting techniques used to detect a specific DNA, RNA or protein molecule in a sample. ![]() The main difference between Southern Northern and Western blotting is that the Southern blotting involves the identification of DNA, and the Northern blotting involves the identification of RNA, whereas the Western blotting involves the identification of proteins.
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