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Freeze-drying protective agent and application thereof in nucleic acid amplification reagent

A technology for freeze-drying protective agent and nucleic acid, applied in the field of molecular biology, can solve problems such as large freeze-drying volume, and achieve the effects of improving production efficiency, good application prospects, and improving transportation and shelf life.

Inactive Publication Date: 2020-09-15
诺迦(杭州)生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above-mentioned patents generally have the disadvantage of large freeze-drying volume, and there is still a lot of room for improvement in terms of shortening the time-consuming process of the freeze-drying protective agent.

Method used

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  • Freeze-drying protective agent and application thereof in nucleic acid amplification reagent
  • Freeze-drying protective agent and application thereof in nucleic acid amplification reagent
  • Freeze-drying protective agent and application thereof in nucleic acid amplification reagent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Lyoprotectant component test, the operation steps are as follows:

[0027] 1. Preparation of lyophilized preservation solution: prepare 7 groups of lyoprotectants according to Table 1, and use ddH 2 O was adjusted to 100mL.

[0028] Table 1 Formulation of lyoprotectant components

[0029] marshalling Trehalose Mannitol polyethylene glycol 8000 β-cyclodextrin 1 40g 4g 2g 1g 2 40g 2g 2g 1g 3 40g - 2g 1g 4 40g 4g 1g 1g 5 40g 4g - 1g 6 40g 4g 2g 2g 7 40g 4g 2g -

[0030] In Table 1, trehalose, mannitol, polyethylene glycol 8000, and β-cyclodextrin were purchased from Sangon Bioengineering (Shanghai) Co., Ltd. ddH 2 O was purchased from Thermo Fisher Scientific.

[0031] 2. Preparation of nucleic acid amplification reagents: Prepare nucleic acid amplification reagents according to Table 2, and the quantity is 150 for testing.

[0032] Table 2 Nucleic Acid Amplification Reagent Preparation Table ...

Embodiment 2

[0062] To optimize the freeze-drying process parameters, the operation steps are as follows:

[0063] 1. Prepare a lyophilized preservation solution according to the following formula: trehalose 40g, mannitol 4g, polyethylene glycol 80002g, β-cyclodextrin 1g, ddH2O to 100mL.

[0064] 2. Prepare nucleic acid amplification reagents at working concentrations according to Table 2, and the quantity is 60 for testing.

[0065] 3. Mix the nucleic acid amplification reagent prepared according to Table 2 and the lyoprotectant prepared in step 1 of Example 2 evenly at a volume ratio of 4:1 to obtain a nucleic acid amplification system mixture. The nucleic acid amplification system mixture was dispensed into PCR tubes at 5 μL / tube, and divided into 4 groups on average, with 12 PCR tubes in each group.

[0066] The above four groups of reagents were freeze-dried respectively, and the freeze-drying procedures included the pre-freezing stage, the sublimation drying stage and the analytical...

Embodiment 3

[0084] Simulated transportation test at 37°C, the operation steps are as follows:

[0085] 1. Preparation of freeze-dried storage solution: trehalose 40g, mannitol 4g, polyethylene glycol 80002g, β-cyclodextrin 1g, ddH2O to 100mL.

[0086] 2. Prepare nucleic acid amplification reagents at working concentrations according to Table 2, and the quantity is 200 for testing.

[0087] 3. Mix the nucleic acid amplification reagent prepared according to Table 2 and the lyoprotectant prepared in step 1 of Example 3 evenly at a volume ratio of 4:1 to obtain a nucleic acid amplification system mixture. The nucleic acid amplification system mixture was dispensed into PCR tubes at 5 μL / tube, and divided into 10 groups on average, with 12 PCR tubes in each group. The 5 groups of reagents were freeze-dried according to the process parameters of group 4 in Tables 6.1-6.3. The other 5 groups of reagents were temporarily stored in a -20°C freezer.

[0088] 4. Packaging: Quickly take out the f...

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Abstract

The invention discloses a freeze-drying protective agent and application thereof in a nucleic acid amplification reagent. The freeze-drying protective agent contains the following substances, including trehalose, mannitol, [beta]-cyclodextrin, polyethylene glycol 8000 and water. The freeze-drying protective agent can obviously improve the transportation and storage life of the nucleic acid amplification freeze-drying reagent under a room temperature condition. In addition, the volume of the reagent is reduced to shorten freeze-drying technical time consumption, production efficiency is improved, and the freeze-drying protective agent has a good application prospect.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a freeze-drying protective agent and its application in nucleic acid amplification reagents. Background technique [0002] Although the development of nucleic acid amplification technology has long been mature and has achieved large-scale industrial application in the field of biomedicine, the transportation and storage of nucleic acid amplification reagents have not been able to get rid of the dependence on the cold chain. Nucleic acid amplification reagents have complex components and contain a variety of biologically active components, and any change in the activity of any component will affect the performance of the reagent. In order to maintain the biological activity of the reagents, most reagent manufacturers first store the key components of the reagents, such as enzyme reagents, separately from other components, and then carry out cold chain transportation and frozen sto...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12Q1/6806
CPCC12N15/10C12Q1/6806C12Q2527/125
Inventor 孙刚朱晓进范春雷李铭夫
Owner 诺迦(杭州)生物工程有限公司
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