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High speed nucleic acid amplification method, and equipment and application thereof

A nucleic acid and extremely fast technology, applied in biochemical equipment and methods, specific-purpose bioreactors/fermenters, microbial measurement/inspection, etc., can solve the problem of low efficiency of PCR instruments, affecting work efficiency, and long reaction time and other issues, to achieve maximum flexibility and applicability, save time, and achieve the effect of rapid nucleic acid detection

Active Publication Date: 2019-08-09
GUANGZHOU ANGEL BIOSAFETY TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because the current PCR process still has a long reaction time and needs batch reactions, that is, the problem of a long cycle, the next batch of reactions must wait for the completion of the previous batch of reactions, which causes the PCR instrument to be inefficiently used. It also affects work efficiency

Method used

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  • High speed nucleic acid amplification method, and equipment and application thereof
  • High speed nucleic acid amplification method, and equipment and application thereof
  • High speed nucleic acid amplification method, and equipment and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Preliminary test on the influence of high temperature ambient temperature on the temperature change rate of heating

[0027] Carry out a preliminary test of the influence of ambient temperature on the temperature change efficiency, the specific operation is as follows:

[0028] Set the temperature of the reaction well of the PCR instrument to 100°C, and use a spot thermometer to test the initial temperature of the liquid in the PCR reaction tube as 60°C. Place the reaction tube in the reaction well of the PCR instrument. When the temperature of the liquid in the PCR reaction tube reaches At 92°C, it takes 14 seconds.

[0029] Set the temperature of the reaction well of the PCR instrument to 95°C, and use a spot thermometer to test the initial temperature of the liquid in the PCR reaction tube as 60°C, place the reaction tube in the reaction well of the PCR instrument, and when the temperature of the liquid in the PCR reaction tube reaches At 92°C, it takes 22...

Embodiment 2

[0034] Example 2 Preliminary test on the influence of low temperature ambient temperature on the temperature change rate of cooling

[0035] Carry out a preliminary test of the influence of ambient temperature on the temperature change efficiency, the specific operation is as follows:

[0036] Set the temperature of the reaction well of the PCR instrument to 10°C, and use a spot thermometer to test the initial temperature of the liquid in the PCR reaction tube as 92°C. Place the reaction tube in the reaction well of the PCR instrument. When the temperature of the liquid in the PCR reaction tube drops It takes 4 seconds to reach 60°C.

[0037] Set the temperature of the reaction well of the PCR instrument to 20°C, and use a spot thermometer to test the initial temperature of the liquid in the PCR reaction tube as 92°C, place the reaction tube in the reaction well of the PCR instrument, and when the temperature of the liquid in the PCR reaction tube drops It takes 5 seconds to ...

Embodiment 3

[0043] A preferred embodiment of the comparative experiment of embodiment 3 extremely fast PCR method and routine PCR method

[0044] Four groups of different short fragments of the hepatitis B virus (HBV) gene were used as the target fragments for amplification, and the ultra-fast PCR method was compared with the conventional PCR method.

[0045] 1. The four sets of target amplification sequences and the corresponding forward primer and reverse primer sequences are as follows:

[0046] The first group of target amplification sequences SEQ ID NO: 1:

[0047] 5'-CCCCAACCTCCAATCACTCACCAACCTCTTGTCCTCCAATTTGTCCTGGTTATCGCTGGATGTGTCTGCG-3'

[0048] The first set of forward primers SEQ ID NO:2: 5'-CCCCAACCTCCAATCACTCA-3'

[0049] The first set of reverse primers SEQ ID NO: 3: 5'-CGCAGACACATCCAGCGATAAC-3'

[0050] The second group of target amplification sequences SEQ ID NO:4:

[0051] 5’-CCCCAACCTCCAATCACTCACCAACCTCTTGTCCTCCAATTTGTCCTGGTTATCGCTGGATGTGTCTGCGGCGTTTTATCATCCTTCCTCTTC...

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Abstract

The invention discloses a high speed nucleic acid amplification detection method. A nucleic acid amplification reaction tube is repeatedly placed in an ultrahigh temperature zone and an ultralow temperature zone for a short period; the temperature of the ultrahigh temperature zone is in a range of 100 to 150 DEG C; the temperature of the ultralow temperature zone is in a range of 10 to 40 DEG C; after 30 to 45 cycles, and the nucleic acid amplification detection is completed within 8 to 15 minutes. The invention also discloses a reactor for the high speed nucleic acid amplification, which comprises an ultrahigh temperature reaction zone with a fixed temperature in a range of 100 to 150 DEG C and an ultralow temperature reaction zone with a fixed temperature in a range of 10 to 40 DEG C. The invention also discloses an operation software system and equipment for the high speed nucleic acid amplification.

Description

technical field [0001] The present invention relates to a gene detection method used in the fields of biology, medicine, etc., and its equipment and application, in particular to a method for extremely fast nucleic acid amplification, and its equipment and application. Background technique [0002] Polymerase Chain Reaction (Polymerase Chain Reaction), referred to as PCR, has become a routine technique for nucleic acid detection. A typical PCR program usually consists of three basic reaction steps of repeated high temperature denaturation-low temperature renaturation-suitable temperature extension, that is, high temperature treatment at 90-95°C for about 10-30 seconds, and then cooling down to 50-65°C, maintain for about 10-30 seconds, then raise the temperature to about 72°C, maintain for 30-60 seconds, and so on for 30-40 cycles. According to the typical PCR three-step cycle amplification process program recorded in the book "Molecular Cloning Experiment Guide", it is usu...

Claims

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Application Information

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IPC IPC(8): C12Q1/6806C12M1/38C12M1/00
CPCC12Q1/6806
Inventor 周荣苏晓波高文娟李潇刘文宽周银华周志超许多
Owner GUANGZHOU ANGEL BIOSAFETY TECH CO LTD
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