Standard sample-based real-time fluorescence PCR relative calibration method

A technology of real-time fluorescence and calibration method, applied in the intersection of optics and biotechnology, can solve the problems of low sensitivity, interference of fluorescence signal, influence of detection results, etc., and achieve the effect of high precision

Inactive Publication Date: 2011-02-23
CHINA JILIANG UNIV
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Problems solved by technology

[0003] At present, most of the real-time fluorescent PCR instruments use CCD as the detector. Its biggest advantage is that it can scan multiple fluorescent signals at the same time. great influence

Method used

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  • Standard sample-based real-time fluorescence PCR relative calibration method
  • Standard sample-based real-time fluorescence PCR relative calibration method
  • Standard sample-based real-time fluorescence PCR relative calibration method

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Embodiment Construction

[0032] The hardware required in the method of the present invention includes: a calibrated fluorescence detector, a computer, a real-time fluorescent PCR instrument to be calibrated and a printer. The calibrated fluorescence detector is an instrument calibrated by the National Institute of Metrology, and its accuracy is higher than that of the general real-time fluorescent PCR instrument, which can be used as standard data; the excitation light source adopts monochrome LED and integrating sphere, and the photodetector is a photomultiplier tube , can detect the standard fluorescent light intensity of fluorescent substances, and the detected data can be saved in the computer through the RS232 serial port of the instrument control box.

[0033] The photodetector of the real-time fluorescent PCR instrument to be calibrated can be a CCD or a photomultiplier tube. When the sample test tube is placed in the instrument, the instrument is normally started to run the entire cycle, and th...

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Abstract

The invention relates to standard sample-based real-time fluorescence polymerase chain reaction (PCR) relative calibration method. In the prior art, no real-time fluorescence PCR calibration method is provided. The method of the invention comprises the following steps of: preparing a standard sample, calculating a calibration standard value, detecting data of an instrument to be calibrated, and determining a calibration result. In the method, fluorescent powder which is divided equally according to a measurement range is adopted so that the fluorescence intensity generated by a fluorescent sample can cover the overall measurement range.

Description

technical field [0001] The invention belongs to the intersection field of optics and biotechnology, and relates to a calibration method of real-time fluorescent PCR, in particular to a relative calibration method of real-time fluorescent PCR based on a standard sample. Background technique [0002] Polymerase Chain Reaction (PCR) technology is a nucleic acid amplification technology that simulates the natural DNA replication process in vitro, also known as cell-free molecular cloning technology. Its principle is similar to the replication of natural DNA and is an in vitro enzymatic reaction. A method of selectively synthesizing specific DNA. Real-time fluorescent PCR technology was launched by Applied biosystems (ABI) in 1996. It is a method of adding fluorescent groups to the PCR reaction system, using real-time detection of fluorescent signal accumulation to detect the entire PCR process, and finally through the calibration curve. Unknown templates were quantified. This ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10G01N21/64
Inventor 程琦孔明叶子弘俞晓平崔海峰
Owner CHINA JILIANG UNIV
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