Medicilon in Situ Hybridization Technology Service

Definition: in situ hybridization (insituhybridization) is a kind of molecular hybridization, which is a kind of hybridization of the labeled probe directly to the nucleic acid to be tested in the tissue or cell, and the principle of immunocytochemistry uses a suitable detection system to target A method for precise and quantitative positioning of nucleic acid sequences.

In situ hybridization technology service

In situ hybridization
A technology developed on the basis of studying the principles of DNA molecular replication. The basic principle is that two single-stranded nucleotide fragments, under suitable conditions, can be combined by hydrogen bonding to form the characteristics of DNA-DNA, DNA-RNA or RNA-RNA double bond molecules, which should be labeled (with Radioactive isotopes, such as 3H, 35S, 32P, fluorescein biotin, digoxin and other non-radioactive substances) DAN or RNA fragments are used as nucleic acid probes, and the nucleic acid to be tested in tissue sections or cells. (RNA or DNA) fragments are hybridized, and then can be displayed by autoradiography and other methods. Observe the presence and location of target mRNA or DAN under light or electron microscope; in situ hybridization can be used to study the synthesis of certain cells in situ Gene expression of polypeptides or proteins. This method has high sensitivity and specificity, and can further explore the functional expression and regulation mechanism of cells from molecular water. It has become an important means of cell biology and molecular biology research.

Development process: Pardue and Gall first developed this technology in 1969. At that time, the only marker available was a radioisotope, and the autoradiography technology obviously became a method of probe sequence detection. However, the development and application of in situ hybridization technology is greatly restricted due to factors such as strict safety requirements, short validity period, long imaging time, and low resolution caused by radioactive decay. Until the emergence of non-radioactive labeled probes, all obstacles encountered in in situ hybridization were completely eliminated, making the technology widely used.

Method: Non-radioactive in situ hybridization technology is divided into two types according to the type of labeled probe. Direct detection method (using fluorescein to directly label the probe) and indirect detection method (using digoxigenin or biotin-labeled nucleotides as a substrate and synthesizing the labeled probe by PCR.)

Classification of in situ hybridization techniques

① Genome in situ hybridization technology: It mainly uses the differences in DNA homology between species, and uses the genomic DNA of another species as blocking at an appropriate concentration to perform in situ hybridization on the target chromosome.
②Fluorescence in situ hybridization technology: use fluorescently labeled nucleic acid fragments as probes to hybridize with specific fl-N:~ on chromosomes or DNA microsections, and detect the signal DNA sequence on the chromosome through a fluorescence detection system (fluorescence microscope) Or the target DNA sequence on the DNA microsection to determine the hybridization site.
③Multi-color fluorescence in situ hybridization technology: It is a new technology developed on the basis of fluorescence in situ hybridization technology. It uses several different colors of fluorescein individually or mixed-labeled probes for in situ hybridization, which can simultaneously Multiple targets are detected, and the colors of each target are different under the fluorescence microscope and on the photo, showing multiple colors, so it is called multi-color fluorescence in situ hybridization.
④In situ PCR: It is the organic combination of conventional in situ hybridization technology and PCR technology, that is, the target nucleic acid sequence is amplified in situ on the chromosome or in the tissue cell through PCR to increase the copy number, and then through in situ hybridization Technology for detection, so as to qualitatively, locate and quantitatively analyze the target nucleic acid sequence. In situ PCR technology has greatly improved the sensitivity and specificity of in situ hybridization technology, and can be used for low-copy or even single-copy gene mapping, providing a broader development prospect for the development of in situ hybridization technology.

The above is the relevant introduction about in situ hybridization, the content comes from the official website of Medicilon.