Extraction method of acidophilic bacteria genome DNA in wet method metallurgy

A technology of hydrometallurgy and extraction method, which is applied in the field of extraction of acidophilic bacteria genome DNA, which can solve the problems of cumbersome procedures, inapplicability of acidophilic bacteria, and unsatisfactory results

Inactive Publication Date: 2007-07-04
RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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Problems solved by technology

However, the acidophilic bacteria in hydrometallurgy exist in the slurry whose pH is lower than 2.0 in the reactor, and the existence state is very special, so the methods used to extract the genomic DNA of other microorganisms are not suitable for acidophilic bacteria
The procedures for DNA extraction of acidophilus reported in relevant literature are cumbersome and the results are not satisfactory

Method used

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  • Extraction method of acidophilic bacteria genome DNA in wet method metallurgy

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Experimental program
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Embodiment 1

[0016] Add sulfur powder to the leatheren medium to a final concentration of 2%, adjust the pH to 2.0 with sulfuric acid, inoculate 10% acidophilus, 170rpm, 40°C constant temperature shaking culture for 48 hours. The bacteria were collected by centrifugation, and the extraction results of the genomic DNA were shown in the second lane of the attached drawing, and the yield of the extracted DNA was 35 μg / g pulp.

Embodiment 2

[0018] Take a high-arsenic refractory gold ore, prepare pulp with 9K medium, pulp concentration 15%, pH 1.8, inoculate acidophilus 10%, 170rpm, 40°C constant temperature shaking culture for 48 hours. The bacteria were collected by centrifugation, and the extraction results of the genomic DNA are shown in the third lane of the attached drawing, and the yield of the extracted DNA was 45 μg / g pulp.

Embodiment 3

[0020] Take a sphalerite concentrate, prepare pulp with 9K medium, pulp concentration 10%, pH 2.0, inoculate 10% acidophilus, 170rpm, 40°C constant temperature shaking culture for 48 hours. The bacteria were collected by centrifugation, and the extraction results of the genomic DNA were shown in the third lane of the attached drawing, and the yield of the extracted DNA was 50 μg / g pulp.

[0021] use A 260 / A 280 The quality of the three genomic DNAs extracted above was tested, and the ratios all reached 1.8, and the results showed that the purity of the three DNAs was relatively high.

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Abstract

The invention provides a novel, fast and stable method for extracting genome DNA from pulp acidophilic bacteria. It comprises following steps: centrifuging cultured pulp, washing with 0.2 M sulfate and SET buffer solution respectively; adding lysozyme and treating for 30 minutes at 37 Deg. C, adding protein enzyme K and SDS and treating for 45 minutes at 50 Deg. C, adding glass balls and vortexing for 1 minute to crack cell completely, then extrcting protein with chloroform, washing with waterless alcohol precipitation and alcohol of 75% and getting pure genome DNA, which can be used for PCR gene operation.

Description

technical field [0001] The invention relates to a method for extracting acidophilic bacteria genome DNA. technical background [0002] The method of using acidophilus to dissolve and recover metals from lean ore and tailings is called biohydrometallurgy. Copper recovery from mine leach water has been known since the 18th century at the Rio Tinto mines in Spain. Biohydrometallurgy was officially recognized as a technology in the 1950s after the isolation and identification of Thiobacillus ferrooxidans. In 1958, the United States began to use biohydrometallurgy to extract copper. In 1966, Canada began to extract uranium, and then wet extraction of zinc, cobalt, nickel, gold and other metals was also successful. Biohydrometallurgy is widely used all over the world. In 1997 and 2001, my country also built two biological copper heap leaching plants with a capacity of more than 1,000 tons in the Dexing Copper Mine in Jiangxi and the Zijinshan Copper Mine in Fujian. In the past ...

Claims

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Application Information

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IPC IPC(8): C12N15/10C22B3/18
CPCY02P10/20
Inventor 张洪勋郝春博白志辉谢慧君张保国丁杰平
Owner RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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