Optimization of the Method for Simultaneous Extraction Bioleaching Bacteria DNA and RNA

Jian Ping Xie; Hui Yun; Guan Zhou Qiu; Xin Xing Liu

doi:10.4028/www.scientific.net/AMR.825.177

Paper Titles

Analysis of Gene Expression in Response to Copper Stress in Acidithiobacillus ferrooxidans Strain D2, Isolated from a Copper Bioleaching Operation
p.157

Temporal Dynamics of Genes Involved in Metabolic Pathways of C and N of L. ferriphilum, in the Industrial Bioleaching Process of Escondida Mine, Chile
p.162

Analysis of Gene Expression as Marker of Relevant Metabolisms, in Three Acidithiobacillus ferrooxidans Strains, in Different Growth Conditions
p.166

Variance Calculations for Quantitative Real-Time PCR Experiments with Multiple Levels of Replication
p.172

Optimization of the Method for Simultaneous Extraction Bioleaching Bacteria DNA and RNA
p.177

Optimization of the Separation Method for Bioleaching Bacteria DNA and RNA Extracted Simultaneously
p.182

How the RegBA Redox Responding System Controls Iron and Sulfur Oxidation in Acidithiobacillus ferrooxidans
p.186

Analysis of the Bacterial Sulphur System
p.190

Heterodisulfide Reductase from Acidithiobacilli is a Key Component Involved in Metabolism of Reduced Inorganic Sulfur Compounds
p.194

HomeAdvanced Materials ResearchAdvanced Materials Research Vol. 825Optimization of the Method for Simultaneous...

Optimization of the Method for Simultaneous Extraction Bioleaching Bacteria DNA and RNA

Abstract:

Metagenomic and metatrascription technologies were developed rapidly and applied widely in environmental microbiology in recent years and were also gradually applied in biohydrometallurgy. However, how to get good quality DNA and RNA simultaneously is still the key factor to determine the success and relibility of the whole data analysis. In this study, a wide used DNA extraction method [ was optimized for extraction of crude nucleic acid from acidic habitat. Our results showed that nucleic acid extraction buffer (pH 7.0) contain PIPES salt could effectively extract crude nucleic acid (including DNA and RNA) from four pure culture and several acidic environmental samples. Qiagen DNA and RNA isolation system (All prep DNA/RNA Mini kit) could separate DNA and RNA. Good quality with A_260/280 was around 1.87±0.01 for total 33.88±7.85 μg DNA and A_260/230 around 2.07±0.07 for total 20.00±2.73 μg RNA were obtained from 300 mL A.f culture medium (strain concentration was around 2×10⁷). All isolated DNA and RNA could be used for downstream molecular analysis. This study provides a useful method for simultaneous extraction, separation and purification DNA and RNA from acidic habitat, which could accelerate the quantitative analysis for biohydrometually and therefore increase the efficiency of biohydrometually.

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Advanced Materials Research (Volume 825)

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177-181

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https://doi.org/10.4028/www.scientific.net/AMR.825.177

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Online since:

October 2013

Authors:

Jian Ping Xie, Hui Yun, Guan Zhou Qiu, Xin Xing Liu

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Biohydrometually, Metagenomic, Metatrascription, Simultaneous Extraction DNA and RNA

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