Method for identifying pseudo lysogenic phages SNJ1 of halophilic archaea

A technology for the identification of halophilic archaea and its identification method, which is applied in the field of identification of pseudolysogenic phage SNJ1 of halophilic archaea, which can solve the problems of high cost, complicated identification process, and long identification period, and achieve reduced workload, The effect of simplifying the identification steps and shortening the identification period

Inactive Publication Date: 2014-04-02
WUHAN POLYTECHNIC UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to overcome the problems of long identification period, complex identification process and high cost in the prior art, and provide a pseudo-halophilic archaea with short identification period, simple operation and low cost. Identification method of lysogenic phage SNJ1

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] A method for identifying pseudolysogenic phage SNJ1 of halophilic archaea, the method comprising the following steps in sequence:

[0040] Step 1: Configuration of medium No. 1 and medium No. 2:

[0041] The configuration method of the No. 1 medium is: first weigh NaCl250g, MgCl 2 ·6H 2 O30g, yeast powder 2g, whey protein hydrolyzate 2.5g, agar 10 ~ 15g, add water and mix to 1000mL to form a solid-liquid mixture, and then the solid-liquid mixture in 0.69 × 10 5 Pa, sterilize at 100-125°C for 10-60 minutes to obtain No. 1 medium;

[0042] The configuration method of the No. 2 medium is: first weigh NaCl250g, MgCl 2 ·6H 2 030g, yeast powder 2g, whey protein hydrolyzate 2.5g, agar 15g and add water and mix and be settled to 1000mL to form solid-liquid mixture, then this solid-liquid mixture is in 0.69 * 10 5 Pa, sterilize at 100-125°C for 10-60 minutes to obtain No. 2 medium;

[0043] Step 2: Cultivation of the halophilic archaea to be tested: first pour No. 1 medium...

Embodiment 2

[0048] The steps are basically the same as in Example 1, the difference is that:

[0049] In step 1, the configuration method of No. 2 medium is: first weigh 250g of NaCl, MgCl 2 ·6H 2 030g, yeast powder 2g, whey protein hydrolyzate 2.5g, agar 4g and add water and mix and be settled to 1000mL to form solid-liquid mixture, then this solid-liquid mixture is in 0.69 * 10 5 Pa, sterilize at 100-125°C for 10-60 minutes to obtain No. 2 medium.

[0050] The detection rate of this implementation method is 96%.

Embodiment 3

[0052] The steps are basically the same as in Example 1, the difference is that:

[0053] In step 4, the inoculation refers to dividing a plurality of equally divided blocks on the back of the plate, then picking a single colony and adding it to 20 μL of liquid medium to mix evenly, and then taking 6 μL of liquid medium mixed with a single colony to drop Add to the equal division blocks on the double-layer plate, wherein, the configuration method of the liquid medium is: first weigh NaCl250g, MgCl 2 ·6H 2 030g, yeast powder 2g, whey protein hydrolyzate 2.5g and add water and mix and be settled to 1000mL to form mixed solution, then this mixed solution is in 0.69 * 10 5 Pa, sterilize at 100-125°C for 10-60 minutes to obtain a liquid medium.

[0054] The detection rate of this implementation method is 100%.

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Abstract

The invention discloses a method for identifying pseudo lysogenic phages SNJ1 of halophilic archaea. The method comprises the following steps: at first, applying the halophilic archaea to be detected to a solid culture medium for culturing for 5-7 days to form single colony; then, inoculating the single colony to a dual-layer flat plate with indicator bacteria in an upper layer, and culturing at constant temperature for 2 days; finally, observing whether the phages are formed on the dual-layer flat plate. According to the method, the phages can be identified without enrichment culture of the single colony and separation of the phages from host cells, so that the identification cycle is shortened, the identification process is simplified, and the cost is reduced.

Description

technical field [0001] The invention relates to an identification method of phage, in particular to an identification method of pseudolysogenic phage SNJ1 of halophilic archaea, which is suitable for shortening the identification period, simplifying the identification process and reducing the cost. Background technique [0002] Halophilic archaea live in high-saline (1.5-5M) environments, including some salt lakes and salt factories. In some environments where the salt concentration is close to saturation (about 35% w / v), the halophilic archaea are still It exists at a higher density (about 108cell / mL). [0003] Phage is a general term for bacterial viruses that infect microorganisms such as bacteria, fungi, actinomycetes, or spirochetes. As a type of virus, phage has some unique characteristics of viruses: the individual is small, does not have a complete cell structure, and only contains a single nucleic acid. The phage genome contains many genes, but all known phages use...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70
CPCC12Q1/04
Inventor 梅运军何聪聪胡纯张顺喜黄咏驰
Owner WUHAN POLYTECHNIC UNIVERSITY
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