RESEARCH ART I C L E
Insights intothe dynamics of bacterial communities during chalcopyrite bioleaching
Zhiguo He1,2, Fengling Gao1,2, Jiancun Zhao1,2, Yuehua Hu1,2 & Guanzhou Qiu1,2
1School of Minerals Processing and Bioengineering, Central South University, Changsha, Hunan, China; and 2Key Laboratory of Biometallurgy, Ministry of Education, Changsha, Hunan, China
Correspondence: Zhiguo He, School of
Minerals Processing and Bioengineering,
Central South University, Changsha, Hunan
410083, China. Tel./fax: 186 731 88879815; e-mail: zhighe@gmail.com
Received 19 December 2009; revised 14 April
2010; accepted 17 June 2010.
Final version published online 3 August 2010.
DOI:10.1111/j.1574-6941.2010.00943.x
Editor: Alfons Stams
Keywords
DGGE; …show more content…

Materials and methods
Site description and sample collection
Samples were collected from the Yunfu sulfide mine, in
Guangdong province, China. Owing to its high concentration of sulfur and low concentrations of other elements, the mine had been used mainly to obtain pyrite for manufacturing sulfuric acid since 1988.
The aqueous AMD sample was pH 2.5, around 25.0 1C, and the concentrations of the elements are shown in Table 1. A 10-
L water sample was collected from the sampling position and processed within 24 h of collection. The sample was filtered through a 0.22-mm hyperfiltration membrane with a vacuum pump. The sediments on the membrane were washed twice with sterile deionized water. The sediments washed from the filtration membrane were then stored at 70 1C. The filtered water sample was prepared for chemical analysis.
DNA extraction and purification
Extraction of nucleic acids was carried out according to the procedure described by Zhou et al. (1996). Using a combination of methods including grinding, freezing, and thawing, and treatment with sodium dodecyl sulfate, various types of bacteria were effectively lysed. The crude DNA was purified using the Wizard plus sv Minipreps DNA purification system (Promega Corporation) and quantified by ethidium bromide-UV detection on an agarose gel.
PCR and