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Freeze-drying protective agent and freeze-drying method of RNA amplification reaction reagent

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

Active Publication Date: 2020-08-21
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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  • Freeze-drying protective agent and freeze-drying method of RNA amplification reaction reagent
  • Freeze-drying protective agent and freeze-drying method of RNA amplification reaction reagent
  • Freeze-drying protective agent and freeze-drying method of RNA amplification reaction reagent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Embodiment 1 A kind of preparation method of RNA amplification reaction microsphere

[0047] Reagent: (500 μL volume)

[0048]

[0049] The preparation method comprises the following steps:

[0050] (1) Preparation of lyoprotectant: take trehalose, mannitol, bovine serum albumin, surfactant, defoamer, lentinan and sucrose in a container and mix evenly;

[0051] (2) Prepare reaction reagents: place buffer, primers, probes and dNTPs in a container and mix them evenly;

[0052] (3) mix the lyoprotectant prepared in step (1) with the reaction reagent prepared in step (2) and then add the enzyme mixture to prepare a lyophilized reagent;

[0053] (4) Use a pipette gun to absorb the lyophilized reagent prepared in step (3), and then drop it drop by drop into liquid nitrogen for lyophilization to make microspheres, and transfer the lyophilized microspheres to the pre-frozen freezer Continue freeze-drying in the drier to obtain the finished product of freeze-dried microsph...

Embodiment 2

[0056] Embodiment 2 A kind of preparation method of RNA amplification reaction microsphere

[0057] Reagent: (500 μL volume)

[0058]

[0059]

[0060] The preparation method comprises the following steps:

[0061] (1) Preparation of lyoprotectant: take trehalose, mannitol, bovine serum albumin, surfactant, defoamer, lentinan and sucrose in a container and mix evenly;

[0062] (2) Prepare reverse transcription polymerase chain reaction reagents: place buffer, primers, probes and dNTPs in a container and mix evenly;

[0063] (3) mix the lyoprotectant prepared in step (1) with the reaction reagent prepared in step (2) and then add the enzyme mixture to prepare a lyophilized reagent;

[0064] (4) Use a pipette gun to absorb the lyophilized reagent prepared in step (3), and then drop it drop by drop into liquid nitrogen for lyophilization to make microspheres, and transfer the lyophilized microspheres to the pre-frozen freezer Continue freeze-drying in the drier to obtain...

Embodiment 3

[0067] Embodiment 3 A kind of preparation method of RNA amplification reaction microsphere

[0068] Reagent: (500 μL volume)

[0069]

[0070]

[0071] The preparation method comprises the following steps:

[0072] (1) Preparation of lyoprotectant: take trehalose, mannitol, bovine serum albumin, surfactant, defoamer, lentinan and sucrose in a container and mix evenly;

[0073] (2) Prepare reverse transcription polymerase chain reaction reagent: mix buffer, primer, probe, dNTPs and enzyme evenly;

[0074] (3) mix the lyoprotectant prepared in step (1) with the reaction reagent prepared in step (2) and then add the enzyme mixture to prepare a lyophilized reagent;

[0075] (4) Use a pipette gun to absorb the lyophilized reagent prepared in step (3), and then drop it drop by drop into liquid nitrogen for lyophilization to make microspheres, and transfer the lyophilized microspheres to the pre-frozen freezer Continue freeze-drying in the drier to obtain the finished produc...

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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 protectant for RNA amplification reaction reagents and a freeze-drying method. Background technique [0002] Polymerase chain reaction (PCR) was invented by Kary Mullis of Centus Corporation in the United States, and was first reported in Science magazine by Saiki et al. in 1985. It is an in vitro rapid amplification DNA technology developed in recent years. Through PCR, a large number of specific nucleic acids can be easily and quickly obtained from trace biological materials by in vitro amplification, which has high sensitivity and specificity, and can be used in the detection of trace samples in animal quarantine, but conventional PCR only It can perform qualitative and quantitative analysis on the final product of the amplification reaction. With the advancement of technology and the deepening of research, it is necessary to perform quantitative analysis on the pr...

Claims

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Application Information

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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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