Last Updated on
August 28, 2023
Blotting techniques are one of the commonly used techniques in molecular biology and life sciences labs to detect and study different molecules, such as proteins and nucleic acids. The term “blotting” refers to the transfer of biomolecules to a membrane.
Based on the molecules separated, such as DNA fragments, RNA, or proteins, the blotting method has three different types:
During the process, the molecules separated using the gel electrophoresis technique in the gel matrix are transferred to blotting membranes, such as nitrocellulose membranes and PVDF membranes. The transferred bands are visualized using different detection methods.
The blotting techniques have applications in many research studies, including gene expression, hybridization, and identifying target sequences, such as specific DNA sequences or specific RNA sequences.
In this article, we will review more about different types of blotting techniques, their working mechanism, and applications in labs and industries.
All three blotting techniques, western, southern, and northern blotting, separate different types of macromolecules and involve different reagents in the process.
Western blotting technique is also known as immunoblotting. The method was developed by Towbin and his colleagues in 1979 to study specific proteins.
The protein blotting procedure involves four stages:
The secondary antibody is conjugated either to an enzyme, fluorescent proteins, or biotin that forms a color change or fluorescence when a suitable substrate is added to the membrane.
Southern Blotting techniques as introduced by Edwin Southern in 1975. The technique detects and analyzes a range of DNA fragments or DNA sequences in a whole DNA sample. The steps involved in the technique include:
The technique is used to detect and analyze specific RNA molecules from a mixed RNA sample. The technique was developed by Alwin in 1979 to study known mRNA sequences. The steps involved in the blotting procedure include:
Other than these, some other immunoblotting techniques (less commonly used) are also known, such as eastern blotting and southwestern blotting assays.
Here’s a table to help you clearly distinguish all three, western, northern, and southern blotting techniques side by side:
So, while experimenting, consider what type of biomolecule you want to analyze using the technique, whether it’s a protein or nucleic acid. This will help you to determine which method is suitable for your lab workflow.
Blotting techniques are methods that involve transferring the separated biomolecules in a gel matrix to the membrane filter, such as nylon, nitrocellulose, or PVDF membrane. The technique is used in many life sciences labs, such as biotechnology and biochemistry for the analysis of a range of biomolecules, such as proteins and nucleic acids.
Three commonly known and extensively utilized blotting techniques include western blotting (for protein analysis), northern blotting (for RNA studies), and southern blotting used for DNA sample analysis.
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