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High-Resolution Comparison of Bacterial Conjugation Frequencies

Article DOI: 10.3791/57812-v • 05:18 min • January 10th, 2019
January 10th, 2019 •
1,2,3, 3, 1
1Applied Chemistry and Biochemical Engineering Course, Department of Engineering, Graduate School of Integrated Science and Technology, Shizuoka University, 2Department of Bioscience, Graduated School of Science and Technology, Shizuoka University, 3Japan Collection of Microorganisms, RIKEN BioResource Research Center

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With an aim to understand the behaviors of various bacterial conjugative DNA elements under different conditions, we describe a protocol for detecting differences in conjugation frequency, with high resolution, to estimate how efficiently the donor bacterium initiates conjugation.

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High-resolution Bacterial Conjugation Frequencies Microbiology Field Plasmid DNA Spread Technique Background Reduction Small Differences High Resolution Procedure Kosuke Yanagiya Graduate Student Calculate Conjugation Frequency Most Probable Number Filter Mate Overnight Donor Culture Overnight Recipient Culture Co-culture Incubation Serial Dilute Original Donor And Recipient Cultures Plating In Triplicate Luria Broth LB Kanamycin Gentamycin Plates Two-day Culture Resuspend The Culture On The Filter Sterile LB Containing Kanamycin And Gentamycin 96-well Cell Culture Plate Quadruplicate Counting Colony-forming Units (CFUs) Transconjugants Growth
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