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Freeze-dried microsphere of PCR reaction agent and preparing method thereof

A technology of reaction reagents and microspheres, which is applied in biochemical equipment and methods, measurement/inspection of microorganisms, etc., can solve the problems that PCR reagents cannot be stored and transported at room temperature, and achieve uniform size and shape, convenient access, and simplified operation steps Effect

Pending Publication Date: 2019-11-15
INTEGRATED BIOSYSTEMS CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide a freeze-dried microsphere of PCR reaction reagent and its preparation method, to solve the problem in the prior art that PCR reagents cannot be stored and transported at room temperature

Method used

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  • Freeze-dried microsphere of PCR reaction agent and preparing method thereof
  • Freeze-dried microsphere of PCR reaction agent and preparing method thereof
  • Freeze-dried microsphere of PCR reaction agent and preparing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment example 1

[0031] 1. Preparation of PCR reaction reagent containing lyoprotectant:

[0032] Table 1

[0033] .

[0034] Replenish water to 250μl volume, invert to mix, briefly centrifuge, and place on ice box for use.

[0035] 2. Freeze-drying of PCR reaction reagents.

[0036] 1. Pour liquid nitrogen into a sterilized container, use a pipette gun that can be accurately quantified, drop the prepared mixed solution into the liquid nitrogen with a volume of 6 μl, condense into a round ball, and wait for 10 seconds Left and right, the ball sank to the bottom of the liquid nitrogen, picked up the ball, and transferred it to a vial that had been pre-cooled with dry ice.

[0037] 2. After the reagent pellets are collected, transfer them to a freeze dryer. The freeze-drying procedure is as follows:

[0038] Table 2

[0039] .

[0040] 3. After freeze-drying, the freeze-dried microspheres of PCR reaction reagents are obtained, see figure 1 .

[0041] 3. Observing the freeze-dried mic...

Embodiment example 2

[0057] 1. Preparation of PCR reaction reagent containing lyoprotectant:

[0058] The preparation method is shown in Table 1 in Example 1.

[0059] 2. Freeze-drying of PCR reaction reagents.

[0060] 1. Pour liquid nitrogen into a sterilized container, use a pipette gun that can be accurately quantified, drop the prepared mixed solution into the liquid nitrogen with a volume of 6 μl, condense into a round ball, and wait for 10 seconds Left and right, the ball sank to the bottom of the liquid nitrogen, picked up the ball, and transferred it to a vial that had been pre-cooled with dry ice.

[0061] 2. After the reagent pellets are collected, transfer them to a lyophilizer for vacuum freeze-drying:

[0062] The freeze-drying procedure is shown in Table 2 in Example 1.

[0063] 3. After freeze-drying, the PCR reaction reagent freeze-dried microspheres are obtained.

[0064] 3. Observe the freeze-dried microspheres under a microscope and measure their diameter:

[0065] Table 4 ...

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Abstract

The invention provides a freeze-dried microsphere of a PCR reaction agent. The freeze-dried microsphere comprises an enzyme reaction liquid, desoxyribonucleic acid triphosphoric acid, a primer, a template, a biological buffer solution, glucan, trehalose, bovine serum albumin, gelatin, glycerinum, dimethyl sulfoxide, a surfactant, a defoaming agent, a preservative and the like. By means of the freeze-dried microsphere of the PCR reaction agent and the preparing method thereof, the problem that a PCR agent cannot be preserved or transported at a normal temperature in the prior art is solved; theproblems that the PCR agent can hardly be packaged or stored and a freeze-dried powder agent can hardly be quantified are solved. The freeze-dried product comprises a PCR reaction main agent and is convenient to use and saves time, and the contamination introduced in the experiment operation process and the errors of reducing the addition of reaction components are effectively avoided.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a freeze-dried microsphere of a PCR reaction reagent and a preparation method thereof. Background technique [0002] Polymerase Chain Reaction (PCR) uses DNA to denature into a single strand at high temperature (often around 95°C) in vitro, and primers and single strands are paired according to base complementarity at low temperature (often around 60°C) Combining with the principles, adjust the temperature to the optimal reaction temperature of DNA polymerase (about 72°C), and DNA polymerase synthesizes complementary strands along the direction from phosphoric acid to five-carbon sugar (5'-3'). PCR was invented by Mullis of the American PE company in 1985. It has the outstanding advantages of specificity, sensitivity, high yield, rapidity, simplicity, good repeatability, and easy automation. [0003] In recent years, PCR reagents have been widely used in food testing, pathogen detec...

Claims

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Application Information

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IPC IPC(8): C12Q1/6806C12Q1/686
CPCC12Q1/6806C12Q1/686C12Q2563/149
Inventor 代有来高静蔡亦梅张瑜范东雨李洁昆任鲁风
Owner INTEGRATED BIOSYSTEMS CO LTD
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