Method for labeling nucleic acid DNA with acridine ester
The chemiluminescence reagent acridinium ester is an important detection reagent in CLIA. Its appearance is a yellow powder. Its detection sensitivity is very high. It has gradually become the mainstream detection method in the domestic in vitro diagnostic field. Objects that can be detected and labeled by acridinium esters include proteins, antibodies, nucleic acid DNA or RNA, etc., with a wide range of applications.
In terms of in vitro diagnosis, acridinium ester chemiluminescence reagent is very suitable for labeling DNA strands to make chemiluminescent DNA probes. This article will introduce a method for labeling nucleic acid with acridinium ester.
Chemiluminescence reagent acridinium ester
Nucleic acid is the most important substance in all biological molecules. It is widely present in all animal and plant cells and microorganisms. Nucleic acid is divided into two categories: deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). Modern medical research results show that many diseases such as cancer and genetic diseases are related to DNA mutations, and many infectious diseases are caused by viruses, germs or parasites in the environment. Therefore, analysis of virus specific sequence DNA Conducive to the control of the epidemic.
In nucleic acid hybridization analysis, the preparation of labeled probes with strong specificity and high sensitivity is the key to successful nucleic acid hybridization analysis. Acridinium ester derivatives can be directly labeled on nucleic acid probes without the need for catalysts and the luminescence quantum yield is not affected. In addition, under certain conditions, the labeled acridinium ester on the unhybridized single-stranded DNA is hydrolyzed and destroyed, and only the double-stranded protected acridinium ester formed by hybridization can produce chemiluminescence, and the entire hybridization process can be monitored without separation.
This method for labeling nucleic acids with acridinium esters mainly includes three steps. Firstly, the 5'and 3'ends of the DNA probes are protected respectively; then the acridinium ester labeling is carried out, and finally the DNA is purified and separated by HPLC. Among them, acridinium ester labeling is the most important. Dissolve 25mM acridinium ester in dimethyl sulfoxide (DMSO), configure 1M HEPES buffer (PH=8.0), and follow the molar ratio of nucleic acid probe: acridinium ester=1:5 Add to HEPES buffer and react at 37°C for 1h.
This method creatively labeled acridinium esters on both ends of the DNA, further enhancing the sensitivity of detection. Acridine esters have the advantages of mild reaction conditions, good reproducibility, high luminous efficiency, and strong luminous intensity. They are widely used in the fields of inorganic and organic compounds, environmental monitoring, biological and pharmaceutical analysis, and are also commonly used in various types of diseases. Sensitive detection and diagnosis.
Acridine ester chemiluminescence reagents include two types of acridine ester and acridine sulfonamide. There are six acridinium ester reagents developed by Desheng, which can meet the different needs of labeling proteins, antibodies, nucleic acids and other molecules.
MOPS buffer is an important biochemical reagent used to maintain the acid-base balance and ion environment of tissue samples, protecting cell structure and function. Widely used in cell culture, tissue fixation, and immunohistochemical staining to improve experimental accuracy and reliability. Understanding the characteristics and application principles of MOPS buffer is crucial for biomedical research.