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Uses of hydroxypropyl-beta-cyclodextrin in preparation of nucleic acid freeze-drying protection agent

A technology of drying protective agent and hydroxypropyl, applied in the field of hydroxypropyl-β-cyclodextrin, can solve the problems of unfavorable long-distance transportation of nucleic acid, inconvenient experimental operation and the like

Active Publication Date: 2018-05-25
CHINA ANIMAL DISEASE CONTROL CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

On the one hand, it needs to be equipped with complex equipment such as a -70°C refrigerator, on the other hand, it is not conducive to the long-distance transportation of nucleic acids, which brings great inconvenience to the experimental operation

Method used

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  • Uses of hydroxypropyl-beta-cyclodextrin in preparation of nucleic acid freeze-drying protection agent
  • Uses of hydroxypropyl-beta-cyclodextrin in preparation of nucleic acid freeze-drying protection agent
  • Uses of hydroxypropyl-beta-cyclodextrin in preparation of nucleic acid freeze-drying protection agent

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Experimental program
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Embodiment 1

[0032] Example 1. Application of hydroxypropyl-β-cyclodextrin in the preparation of nucleic acid freeze-drying protective agent

[0033] 1. Study on volume change of different concentrations of hydroxypropyl-β-cyclodextrin after freeze-drying

[0034] Take hydroxypropyl-β-cyclodextrin powder, prepare 1mg / mL, 5mg / mL, 10mg / mL, 20mg / mL, 50mg / mL, 100mg / mL and 500mg / mL solutions with ultrapure water, take Put 0.8mL solution into a 1.5mL centrifuge tube, freeze-dry at -20°C, observe the shape of the solid component after freeze-drying, and use 100ng / μL DNA solution (the solvent is TE buffer) to freeze-dry the sample as a control.

[0035]The results showed that the solid powder formed after the direct freeze-drying of the DNA solution (the solvent was TE buffer) could not be observed with the naked eye, while the hydroxypropyl-β-cyclodextrin solution had an obvious solid form after freeze-drying. When the concentration of hydroxypropyl-β-cyclodextrin solution is greater than 5mg / mL...

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Abstract

The invention discloses uses of hydroxypropyl-beta-cyclodextrin in preparation of a nucleic acid freeze-drying protection agent. According to the present invention, DNA can be well protected by usinghydroxypropyl-beta-cyclodextrin or a hydroxypropyl-beta-cyclodextrin aqueous solution as a DNA freeze-drying protection agent, RNA can be well protected by using a composition H and a solution H as RNA freeze-drying protection agents, and nucleic acid quantitative analysis, subsequent DNA digestion, PCR / RT-PCR amplification, fluorescence quantitative PCR / RT-PCR amplification and other operations are not affected; and RNA, hydroxypropyl-beta-cyclodextrin and DEPC are mixed, and the mixture is freeze-dried, wherein the RNA stored within 6 months at the room temperature has the variation coefficient of 5.90%, and is significantly superior to the RNA stored in a -70 DEG C liquid state (the variation coefficient is 13.33%), and the storing condition does not require the arrangement of -70 DEG Cultra-low temperature refrigerator and other complex equipment so as to easily achieve the long-distance transportation and storage of RNA samples.

Description

technical field [0001] The invention relates to the use of hydroxypropyl-β-cyclodextrin in the field of molecular biology, in particular to the use of hydroxypropyl-beta-cyclodextrin in preparing nucleic acid freeze-drying protective agents. Background technique [0002] As one of the most basic substances of life, nucleic acid is an important object of life science research and biological detection, and is also an important component of biological nucleic acid products and nucleic acid reagents. According to its chemical composition, it is divided into ribonucleic acid (RNA) and deoxyribonucleic acid. (DNA) two types. [0003] As a biological macromolecule, nucleic acid is easily degraded under natural conditions. Its stable storage technology plays an important role in the development and production of nucleic acid samples, nucleic acid standard substances, nucleic acid kits, and nucleic acid vaccines. Therefore, advanced nucleic acid preservation technology has broad app...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12Q1/6806
CPCC12Q1/6806C12Q2527/125
Inventor 王传彬于雷刘洋顾小雪刘玉良
Owner CHINA ANIMAL DISEASE CONTROL CENT
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