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Environmental Engineering Research 2009;14(1): 63-67. DOI: https://doi.org/10.4491/eer.2009.14.1.063
Comparative Quantification of LacZ (β-galactosidase) Gene from a Pure Cultured Escherichia coli K-12
Ji-sun Han, and Chang-gyun Kim
Department of Environmental Engineering, Inha University, Incheon, South Korea
Corresponding Author: Chang-gyun Kim ,Tel: +82-32-860-7561, Fax: +82-32-865-1425, Email: cgk@inha@ac.kr
Received: June 14, 2008;  Accepted: January 17, 2009.
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Escherichia coli K-12 (E. coli K-12) is a representative indicator globally used for distinguishing and monitoring dynamic fates of pathogenic microorganisms in the environment. This study investigated how to most critically quantify lacZ (β-galactosidase) gene in E. coli K-12 by two different real-time polymerase chain reaction (real-time PCR) in association with three different DNA extraction practices. Three DNA extractions, i.e., sodium dodecyl sulfate (SDS)/proteinase K, magnetic beads and guanidium thiocyanate (GTC)/silica matrix were each compared for extracting total genomic DNA from E. coli K-12. Among them, GTC/silica matrix and magnetic beads beating similarly worked out to have the highest (22-23 ng/μL) concentration of DNA extracted, but employing SDS/proteinase K had the lowest (10 ng/μL) concentration of DNA retrieved. There were no significant differences in the quantification of the copy numbers of lacZ gene between SYBR Green I qPCR and QProbe-qPCR. However, SYBR Green I qPCR obtained somewhat higher copy number as 1×108 copies. It was decided that GTC/silica matrix extraction or magnetic beads beating in combination with SYBR Green I qPCR can be preferably applied for more effectively quantifying specific gene from a pure culture of microorganism.
Keywords: E. coli K-12 | DNA extraction | Real-time PCR | QProbe-qPCR | SYBR Green I qPCR
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