Nuclease protection assay
Nuclease Protection Assay
Nuclease Protection Assay (pronounced: noo-klee-ase proh-tek-shun as-ey) is a laboratory method used in molecular biology to analyze specific RNA or DNA sequences. The term originates from the words "nuclease," a type of enzyme that cleaves nucleic acids, and "protection assay," referring to a test that identifies protected fragments of nucleic acids.
History
The Nuclease Protection Assay was first developed in the late 20th century as a tool for studying gene expression. The method has since been refined and is now widely used in molecular biology research.
Methodology
The Nuclease Protection Assay involves the use of a nuclease enzyme to digest unprotected nucleic acids. A labeled probe, complementary to the target sequence, is hybridized to the sample. The nuclease then digests all single-stranded nucleic acids, leaving only the probe-target duplexes. These protected fragments are then separated and analyzed, providing information about the presence and quantity of the target sequence.
Applications
Nuclease Protection Assays are used in various fields of biological research. They are particularly useful in studying gene expression, as they can provide quantitative data on specific mRNA transcripts. They can also be used to detect specific DNA sequences, such as those associated with genetic disorders or infectious diseases.
Related Terms
- molecular biology: The branch of biology that deals with the structure and function of the molecules essential for life.
- nuclease: An enzyme that cleaves the chains of nucleotides in nucleic acids into smaller units.
- gene expression: The process by which information from a gene is used to create a functional product, such as a protein.
- mRNA: Messenger RNA, a type of RNA that carries genetic information from the DNA to the ribosome, where it specifies the amino acid sequence of the protein products of gene expression.
- DNA: Deoxyribonucleic acid, the molecule that carries genetic instructions for the development, functioning, growth and reproduction of all known organisms and many viruses.
External links
- Medical encyclopedia article on Nuclease protection assay
- Wikipedia's article - Nuclease protection assay
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