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Method for quick extraction of total RNA from radioresistant bacterium

An anti-radiation bacteria and rapid technology, applied in the field of RNA extraction, can solve the problem of inability to extract total RNA, and achieve the effects of avoiding pollution, shortening operation time, and simplifying the experimental process.

Inactive Publication Date: 2009-02-11
ZHEJIANG SCI-TECH UNIV +1
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Problems solved by technology

These determine that the total RNA in the radiation-resistant bacteria Deinococcus radiodurans cannot be extracted according to the general method of extracting total RNA from bacteria.

Method used

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  • Method for quick extraction of total RNA from radioresistant bacterium
  • Method for quick extraction of total RNA from radioresistant bacterium

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Embodiment Construction

[0019] In order to make the purpose, technical solutions and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with the accompanying drawings and specific embodiments:

[0020] This rapid method for extracting total RNA from the radioresistant bacterium Deinococcus radiodurans comprises the following steps:

[0021] 1. Take 1 mL of the anti-radiation bacteria solution in the exponential growth phase and put it into a 2 mL centrifuge tube, centrifuge at 4,000 rpm for 4 minutes at room temperature, and remove the supernatant. Add 500 μL of cell lysate, mix thoroughly to suspend the bacteria in the lysate, then add 120-200 μL of 50 mg / mL lysozyme, shake and mix, and incubate at 50°C for 15 minutes;

[0022] Note: The supernatant should be cleaned as much as possible. The formula of the cell lysate (pH7-8) is: 10mM Tris-HCl (pH7.5) (tris-hydrochloric acid buffer), 10mM EDTA (pH8.0) (ethylenediaminetetraacet...

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Abstract

The invention relates to a method for rapidly extracting total RNA from radio resistant bacteria, which comprised the following steps: 1) material: exponential phase of radio resistant bacteria (Deinococcus radiodurans) is taken as the material; 2) the total RNA is extracted and schizolysis is carried out: 500mu L of cell lysis solution is added and is whirled evenly; 120-200mu L 50mg / mL of muramidase is added and is heated up to 50 DEG C for heat preservation for 15min; extraction: 1mL of Ezol is added to be degraded for 15min at the room temperature; 200mu L of chloroform is added to be centrifugated, and supernatant fluid is taken out and is added with 1.5-2.5 times of isopropanol precipitation; washing and storage of samples: RNA samples are washed to be added with DEPC water for solution and stored at 80 DEG C below zero; 3) quality test on the RNA: according to the obtained data and electrophoresis result, the obtained total RNA has high purity quotient, high production rate and good integrity. The method has the advantages that the experiment process is rapid, simple and has good repeatability, which is also applicable to the extract of total RNA of the bacteria with complicated cytoderm structure.

Description

technical field [0001] The invention relates to a method for extracting RNA in the field of biotechnology, mainly a method for quickly extracting total RNA from radiation-resistant bacteria, and is suitable for extracting the total RNA of bacteria with complex cell wall structures. Background technique [0002] The radioresistant bacterium Deinococcus radiodurans is a Gram-negative, non-spore-forming bacterium that is extremely resistant to ionizing radiation, desiccation, ultraviolet light, strong oxidants, and some chemical mutagens. Since the discovery of radioresistant bacteria, they have received great attention in the fields of biology, medicine and environmental engineering, and they have been studied from the level of genomics and proteomics. The current research on small RNA is a hot spot in the biological field. How to extract high-quality total RNA is the key to study the radiation-resistant bacteria Deinococcus radiodurans from this aspect. [0003] The current ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/34C07H21/02
Inventor 王星星郭江峰丁先锋徐根明
Owner ZHEJIANG SCI-TECH UNIV
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