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Chapters
- 00:00Title
- 00:07Introduction
- 00:43End Repair of Sheared Genomic DNA
- 02:10Size-Selection of Genomic DNA by Pulsed-Field Gel Electrophoresis (PFGE)
- 04:48DNA-recovery by Gel Extraction and Ligation of Recovered DNA to the Fosmid Cloning Vector
- 10:18Phage-packaging of Vector-insert Ligation Product and Infection of Phage-Resistant Bacteria
- 14:30Plating and Titering
- 16:34Preparation for Colony Picking
- 17:46Colony Picking, Culturing, and Long-Term Storage
- 20:32Conclusion
Construction d'une bibliothèque fosmide avec l'ADN génomique environnementale isolé du continuum profondeur verticale d'un fjord saisonnière hypoxiques est décrite. La bibliothèque clone résultant est capté dans les plaques 384 puits et archivées pour le séquençage et l'aval de criblage fonctionnel par l'application d'un système de picking colonie automatisés.
Tags
Large Insert Environmental Genomic Library ProductionMicrobesCultivation MethodsMetagenomic ApproachesGenetic ContentIndigenous Microbial CommunitiesGenomic DNA LibrariesCloning TechniquesHypoxic FjordDNA Extraction And Purification ProtocolEnd-repaired DNAPulsed-field Gel Electrophoresis (PFGE)Gel ExtractionDNA FragmentsBase Pairs (Kb)Fosmid CopyControl Vector PCC1
