Bioleaching of Gold Ore

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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…
A Geneamp thermocycler (Biometra, T-Grandient, Germany) was used to incubate reactions through an initial denaturation at 94 1C for 2min, followed by 35 cycles of 94 1C for 40 s, 55 1C for 30 s, and 72 1C for 1min, and completed with an extension period of 10min at 72 1C. Products from the amplification reactions of expected size (about 1500 bp) were pooled and purified before ligation. PCR amplification of the archaeal 16S rRNA gene was carried out following the PCR program described above with two different sets of archaea-specific primers, which were as follows: S-D-Arch-0025-a-S-17 (50-CT GGTTGATCCTGCCAG-30) (Robb et al., 1995) or S-D-Arch- 0344-a-S-20 (50-ACGGGGCGCAGCAGGCGCGA-30) (Weisburg et al., 1991) with S--Univ-1517-a-A-21 (50-ACGGC TACCTTGTTACGACTT-30) (Raskin et al., 1994) to yield 1500- or 1120-bp PCR products, respectively. Cloning, RFLP, and sequencing The purified PCR products were ligated into the vector PGEM-T (Promega Corporation), and used to transform DH5a competent host cells. About 120 white colonies were randomly selected from each library. The transformation efficiency was around 5108 CFUmg1 DNA, as determined using an external control provided with the PGEM-T vector system (Promega Corporation). For RFLP determination and sequencing, the inserted fragments were amplified with the vector-specific T7 and SP6 primers. These unpurified PCR products were digested with two restriction

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