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Freeze-drying protective agent, PCR reagent capable of being freeze-dried and application of freeze-drying protective agent

A technology of freeze-drying protective agent and reagent, which is applied in the field of molecular biology, can solve the problem that it cannot be used as a freeze-drying protective agent for PCR reagents, etc., and achieves the effect of convenient storage and transportation.

Pending Publication Date: 2021-01-12
苏州创澜生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these commonly used lyoprotectants have a large degree of inhibition on the PCR reaction itself, so they cannot be used as lyoprotectants for PCR reagents.

Method used

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  • Freeze-drying protective agent, PCR reagent capable of being freeze-dried and application of freeze-drying protective agent
  • Freeze-drying protective agent, PCR reagent capable of being freeze-dried and application of freeze-drying protective agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Embodiment 1: prepare the PCR reagent of different protective agents

[0018] 1) Preparation of PCR reagents using mannitol as a lyoprotectant (mannitol+)

[0019] Measure 20 μL 1M Tris-HCL (pH=8.0), 50 μL 3M KCl, 80 μL 0.5M MgCl 2 , 10 μL 10mMdNTP, 10 μL Mycobacterium tuberculosis-specific primer probe, 5 μL Taq DNA polymerase and 250 μL 20% mannitol, after mixing evenly, supplement 1 mL with ultrapure water to obtain a PCR reagent using mannitol as a protective agent (mannitol+ ).

[0020] 2) Preparation of PCR reagents (salutamide+) using salutamide as a lyoprotectant

[0021] Measure 20 μL 1M Tris-HCL (pH=8.0), 50 μL 3M KCl, 80 μL 0.5M MgCl 2 , 10 μL 10mMdNTP, 10 μL Mycobacterium tuberculosis-specific primer probe, 5 μL Taq DNA polymerase and 500 μL 10% salutamide, after mixing evenly, supplement 1 mL with ultrapure water to obtain a PCR reagent using salutamide as a protective agent (salutamide Lutamine+).

Embodiment 2

[0022] Embodiment 2: Comparison of the impact of lyoprotectants on the performance of PCR reagents

[0023] Take the extracted DNA product of Mycobacterium tuberculosis, detect it with the two PCR reagents prepared in Example 1, and use the PCR reagent without added lyoprotectant as a control.

[0024] Test results such as figure 1 As shown, it can be seen from the results that the use of mannitol as a lyoprotectant has a greater degree of inhibition on the performance of the PCR reagent, while the use of salutamide as a lyoprotectant has almost no effect on the performance of the PCR reagent.

Embodiment 3

[0025] Example 3: Comparison of the effect of using salutamide as the lyoprotectant and the performance of the PCR reagent before and after lyophilization.

[0026] Take the PCR reagent (Salutamide+) prepared in Example 1, and freeze-dry according to the freeze-drying conditions in the following table.

[0027] step pre-freeze a sublimation Analytical drying temperature -45℃ -25℃ 30℃ time 3h 8h 5h Vacuum / 1Pa 1Pa

[0028] The extracted Mycobacterium tuberculosis DNA product was taken, and the reagent after freeze-drying was used for detection, and the reagent before freeze-drying was used as a control.

[0029] Test results such as figure 2 As shown, it can be seen from the results that when salutamide is used as the lyoprotectant, the performance of the PCR reagent before and after lyophilization has little change.

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Abstract

The invention discloses a freeze-drying protective agent. The freeze-drying protective agent comprises the component of sallutamide and can be used for a PCR reagent. After the freeze-drying protective agent is added into the PCR reagent, the performance of the reagent is not affected, and the activity is not lost during freeze-drying. The freeze-dried reagent can be stored at normal temperature,and storage and transportation of the PCR reagent are facilitated.

Description

technical field [0001] The invention belongs to the field of molecular biology, and relates to a freeze-drying protective agent, a freeze-dryable PCR reagent and applications thereof. Background technique [0002] Molecular diagnosis is a technology that uses molecular biology methods to detect the structure or expression level of genetic material in patients to make a diagnosis. Common molecular diagnosis methods include PCR, isothermal amplification, sequencing, etc., among which PCR is currently the most widely used molecule. diagnosis method. [0003] Compared with other diagnostic methods, PCR has the advantages of high detection sensitivity, high accuracy, and strong specificity. However, since the PCR reaction relies on the activity of Taq DNA polymerase, the PCR reagents need to be stored and transported in a low temperature environment (such as -20°C), which causes a lot of inconvenience to the use of PCR reagents. Inactivation due to improper storage of reagents ...

Claims

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Application Information

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IPC IPC(8): C12Q1/686
CPCC12Q1/686C12Q2527/125
Inventor 洪冉
Owner 苏州创澜生物科技有限公司
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