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A kind of freeze-drying protection agent and freeze-drying method of RNA amplification reaction reagent

A technology of freeze-drying protective agent and reagent, which is applied in biochemical equipment and methods, determination/inspection of microorganisms, etc., can solve the problems of poor protection effect of freeze-drying protective agent and short storage time of reagents, and improve freeze-drying effect. , the effect of high sensitivity

Active Publication Date: 2021-01-08
INTEGRATED BIOSYSTEMS CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the protection effect of the lyoprotectant prepared by this application is not good, and the storage time of the reagent is relatively short

Method used

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  • A kind of freeze-drying protection agent and freeze-drying method of RNA amplification reaction reagent
  • A kind of freeze-drying protection agent and freeze-drying method of RNA amplification reaction reagent
  • A kind of freeze-drying protection agent and freeze-drying method of RNA amplification reaction reagent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046]Example 1 A method for preparing RNA amplification reaction microspheres

[0047]Reagents: (volume is 500μL)

[0048]

[0049]The preparation method includes the following steps:

[0050](1) Preparation of freeze-dried protective agent: take trehalose, mannitol, bovine serum albumin, surfactant, defoamer, lentinan and sucrose in a container and mix well;

[0051](2) Preparation of reagents: put buffer, primers, probes and dNTPs in a container and mix well;

[0052](3) Mix the lyophilized protective agent prepared in step (1) with the reaction reagent prepared in step (2) uniformly, and then add the enzyme mixture to prepare a lyophilized reagent;

[0053](4) Use a pipette to suck up the lyophilized reagent prepared in step (3), and then drop by drop into liquid nitrogen for lyophilization to form microspheres, and transfer the completely lyophilized microspheres to the pre-frozen frozen The freeze-drying is continued in the dryer to obtain the finished freeze-dried microspheres.

[0054]The freeze-dr...

Embodiment 2

[0056]Example 2 A method for preparing RNA amplification reaction microspheres

[0057]Reagents: (volume is 500μL)

[0058]

[0059]

[0060]The preparation method includes the following steps:

[0061](1) Preparation of freeze-dried protective agent: take trehalose, mannitol, bovine serum albumin, surfactant, defoamer, lentinan and sucrose in a container and mix well;

[0062](2) Preparation of reverse transcription polymerase chain reaction reagents: put the buffer, primers, probes and dNTPs in a container and mix well;

[0063](3) Mix the lyophilized protective agent prepared in step (1) with the reaction reagent prepared in step (2) uniformly, and then add the enzyme mixture to prepare a lyophilized reagent;

[0064](4) Use a pipette to suck up the lyophilized reagent prepared in step (3), and then drop by drop into liquid nitrogen for lyophilization to form microspheres, and transfer the completely lyophilized microspheres to the pre-frozen frozen The freeze-drying is continued in the dryer to obtain ...

Embodiment 3

[0067]Example 3 A method for preparing RNA amplification reaction microspheres

[0068]Reagents: (volume is 500μL)

[0069]

[0070]

[0071]The preparation method includes the following steps:

[0072](1) Preparation of freeze-dried protective agent: take trehalose, mannitol, bovine serum albumin, surfactant, defoamer, lentinan and sucrose in a container and mix well;

[0073](2) Preparation of reverse transcription polymerase chain reaction reagents: mix the buffer, primers, probes, dNTPs and enzymes uniformly;

[0074](3) Mix the lyophilized protective agent prepared in step (1) with the reaction reagent prepared in step (2) uniformly, and then add the enzyme mixture to prepare a lyophilized reagent;

[0075](4) Use a pipette to suck up the lyophilized reagent prepared in step (3), and then drop by drop into liquid nitrogen for lyophilization to form microspheres, and transfer the completely lyophilized microspheres to the pre-frozen frozen The freeze-drying is continued in the dryer to obtain the finis...

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Abstract

The invention discloses a freeze-drying protective agent and a freeze-drying method of an RNA amplification reaction reagent, and relates to the technical field of biology. The freeze-drying protective agent is prepared from the following components: 5 to 20 percent of trehalose, 5 to 15 percent of mannitol, 0.5 to 5mg / mL of bovine serum albumin, 0.05 to 0.5 percent of a surfactant and 0.01 to 0.05 percent of a defoaming agent. In the implementation process, a small amount of lentinan and cane sugar are added into a freeze-dried reagent; it is found that after addition, the freeze-drying effect of the freeze-dried reagent can be effectively improved, the freeze-dried reagent can be stored for a long time under the normal temperature condition by controlling the concentration ratio of trehalose to lentinan to sucrose, and the freeze-driedreagent still has high sensitivity after being stored for 6 months under the room temperature condition.

Description

Technical field[0001]The invention relates to the field of biotechnology, in particular to a freeze-drying protective agent for RNA amplification reaction reagents and a freeze-drying method.Background technique[0002]Polymerase chain reaction (PCR) was invented by Kary Mullis of Centus, USA, and first reported in Science by Saiki et al. in 1985. It is a rapid in vitro DNA amplification technology developed in recent years. PCR can easily and quickly obtain a large amount of specific nucleic acids from trace biological materials by in vitro amplification. It has high sensitivity and specificity and can be used for the detection of trace samples in animal quarantine, but conventional PCR only The final product of the amplification reaction can be qualitatively and quantitatively analyzed. With the advancement of technology and the deepening of research, it is necessary to quantitatively analyze the products of each cycle of the PCR amplification reaction, so fluorescent quantitative P...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6844
CPCC12Q1/6844
Inventor 高静贾欣月蔡亦梅张瑜金鑫浩任鲁风
Owner INTEGRATED BIOSYSTEMS CO LTD
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