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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 of a fosmid library with environmental genomic DNA isolated from the vertical depth continuum of a seasonally hypoxic fjord is described. The resulting clone library is picked into 384-well plates and archived for downstream sequencing and functional screening by the application of an automated colony picking system.
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
