Deoxyribonucleic acid (DNA) fragment amplification and quantitative detection system based on closed reactors

Abstract

The invention relates to a deoxyribonucleic acid (DNA) fragment amplification and quantitative detection system based on closed reactors, belonging to the technical field of molecular biology detection systems. The DNA fragment amplification and quantitative detection system comprises an amplification unit. A group of closed reactors are arranged in the amplification unit in a matching way. An exciting light device and a fluorescence detection device are arranged on one side of each closed reactor in a matching way. A transmission device which is used for providing power to the closed reactors is arranged at the bottom of the amplification unit in a matching way. The transmission device is connected with a motor. Two or more than two independent constant-temperature areas at different temperatures are arranged in the amplification unit in a matching way. By adopting the technique, the thermal response of reagent samples is improved, the polymerase chain reaction process is accelerated and the experimental period is greatly shortened; the degradation of DNA enzymes is reduced, the amplification sensitivity, accuracy and reproducibility of the samples are improved and the reagent detection linearity range is expanded; the pollution risk during experiments is reduced; the optical path is shortened and the sensitivity and the low background of optical detection are improved; and the low-cost multi-sample multi-channel detection is realized.

Description

technical field [0001] The invention belongs to the technical field of molecular biology detection, and specifically relates to a DNA fragment amplification and quantitative detection system based on a closed reactor, which is widely used in the fields of hospital clinic, inspection and epidemic prevention, scientific research and teaching, agriculture, forestry, biochemistry, criminal investigation and the like. Background technique [0002] Polymerase chain reaction (PCR), referred to as PCR technology, is a technology for amplifying specific DNA fragments in vitro. Its working principle is to use the DNA molecule to be amplified as a template, and use a pair of complementary The oligonucleotide fragment is a primer, and under the action of DNA polymerase, it extends along the template strand according to the mechanism of semi-conservative replication until new DNA synthesis is completed. The continuous repetition of this process can make the amount of DNA synthesis expone...

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Problems solved by technology

But the disadvantages are: due to the cold ambient temperature on the upper side of the plastic test tube, evaporation and condensation water vapor often stays and drips, resulting in sudden changes in fluorescence signals and poor detection repeatability

[0011] Since the total time of the amplification experiment needs 1-2 hours, because the DNA polymerase in the reaction system has the characteristics of rapid degradation at high temperature, the traditional PCR experiment is suitable for low concentration DNA fragments (10 -2 The effect of the experiment is not good, and the coefficient of variation of the Ct value detection is usually CV=1.5 or more, which has an adverse effect on early disease diagnosis clinically

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