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DNA Extraction from 0.22 μM Sterivex Filters and Cesium Chloride Density Gradient Centrifugation
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Journal JoVE Biologie
DNA Extraction from 0.22 μM Sterivex Filters and Cesium Chloride Density Gradient Centrifugation

DNA Extraction from 0.22 μM Sterivex Filters and Cesium Chloride Density Gradient Centrifugation

DOI:
10.3791/1352-v

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15:14 min

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September 18, 2009

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, , , ,
1Department of Microbiology and Immunology,University of British Columbia – UBC

Chapitres

  • 00:05Titre
  • 00:08Introduction
  • 00:47Cell Lysis and Digestion with Proteinase K
  • 02:38DNA Extraction
  • 03:56DNA Concentration and Washing
  • 06:22Determination of DNA Quality and Concentration
  • 07:50Cesium Chloride Gradient Centrifugation
  • 14:13Representative Gel to Check DNA Quality and Concentration
  • 14:40Conclusion

Summary

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We describe a method for extraction of high molecular weight genomic DNA from planktonic biomass concentrated on 0.22 μm Sterivex filters, followed by cesium chloride density gradient centrifugation for purification.

Tags

DNA ExtractionHigh Molecular Weight Genomic DNAPlanktonic BiomassSterivex FiltersStorage/lysis Buffer-80°C StorageCesium Chloride Density Gradient CentrifugationIncubation StepsLiberate DNARemove RNAPhenol:Chloroform ExtractionChloroform ExtractionCentrifugationProtein RemovalCell Membrane RemovalAqueous DNA ExtractBuffer ExchangePurificationCesium Chloride Density GradientSample LimitationProtocol TimeCentrifugation EquipmentHybridization Ovens

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