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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
Bouw van een fosmid bibliotheek met milieu-genomisch DNA geïsoleerd uit de verticale diepte continuüm van een seizoensgebonden hypoxische fjord is beschreven. De resulterende kloon bibliotheek is geplukt in de 384-wells platen en gearchiveerd voor downstream-sequencing en functionele screening door de toepassing van een geautomatiseerd kolonie picking systeem.
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
