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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
Construcción de una biblioteca fosmid con el ADN genómico aislado del medio ambiente de la continuidad de profundidad vertical de un fiordo de la temporada hipóxica se describe. La biblioteca clon resultante se recoge en placas de 384 pocillos y se archivan para la secuenciación de aguas abajo y la detección funcional mediante la aplicación de un sistema de recogida automatizada de la colonia.
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
