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Comparative Quantification of LacZ (β-galactosidase) Gene from a Pure Cultured Escherichia coli K-12
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  • Journal title : Environmental Engineering Research
  • Volume 14, Issue 1,  2009, pp.63-67
  • Publisher : Korean Society of Environmental Engineering
  • DOI : 10.4491/eer.2009.14.1.063
 Title & Authors
Comparative Quantification of LacZ (β-galactosidase) Gene from a Pure Cultured Escherichia coli K-12
Han, Ji-Sun; Kim, Chang-Gyun;
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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 (-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/) concentration of DNA extracted, but employing SDS/proteinase K had the lowest (10 ng/) 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 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.
E. coli K-12;DNA extraction;Real-time PCR;QProbe-qPCR;SYBR Green I qPCR;
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