According to the purpose of DNA amplification and the standard of detection, the PCR instrument can be divided into common PCR instrument, gradient PCR instrument, in situ PCR, real-time fluorescence quantitative PCR instrument and the like.
Ordinary PCR instrument
A PCR instrument that can only run a specific annealing temperature in a single PCR amplification is called a normal PCR instrument. If you want to use it for different annealing temperatures, you need to run multiple times. Primarily used as a simple, amplification of the annealing temperature of the gene of interest.
Mainly used in scientific research, teaching, clinical medicine, testing, quarantine, etc.
Gradient PCR instrument
One-time PCR amplification can be set up as a gradient PCR machine with a range of different annealing temperature conditions (typically 12 temperature gradients). Because the different amplified DNA fragments are different in annealing temperature, a series of gradient annealing temperatures are used for amplification, so that one-time PCR amplification can screen out the most suitable annealing temperature for high expression. Amplification. It is mainly used to study the amplification of unknown DNA annealing temperature, which saves time and saves money. It can also be used for ordinary PCR without setting a gradient.
Gradient PCR instruments are widely used in scientific research and teaching institutions.
In situ PCR
(Some brands of PCR instruments have the functions of ordinary PCR, gradient PCR, and in situ PCR, and carry out experimental work for multiple purposes through replacement modules)
It is an intracellular gene amplification instrument for localization analysis of target DNA from cells. Such as the location of the pathogenic gene in the cell or the location of the target gene in the cell. The integrity of the cells or tissues can be maintained, and the PCR reaction system can be infiltrated into tissues and cells, and gene amplification can be performed at the position where the target DNA of the cells is located. Not only can the target DNA be detected, but the position of the target sequence within the cell can also be indicated. It is of great practical value to study the pathogenesis and clinical process and pathological changes of diseases at the molecular and cellular levels.
Real-time PCR
On the basis of the design of the common PCR instrument, a fluorescent signal excitation and acquisition system and a computer analysis processing system are added to form an instrument with a fluorescent quantitative PCR function. The principle of PCR amplification is the same as that of ordinary PCR amplification. The primers added during PCR amplification are labeled with isotopes, fluorescein, etc., and primers and fluorescent probes are used to simultaneously bind to the template for specific amplification. The results of the amplification are transmitted to the computer analysis and processing system through the real-time acquisition signal connection of the fluorescence signal acquisition system, and the quantized real-time result output is obtained.
The real-time PCR instrument has a single channel, two channels and multiple channels. When only one fluorescent probe is used, a single channel is used; when there are multiple fluorescent labels, multiple channels are used. Single-channel can also detect multi-fluorescence markers and target gene expression products, because only one target gene can be detected at a time, and multiple amplifications are required to detect the amount of different target gene fragments. Multi-channel facilitates multiplex PCR and enables the detection of multiple genes of interest at a time.
Real-time fluorescence quantitative PCR is mainly used in clinical medical testing, biomedical research and development, food industry, research institutions and so on.
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