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大型插入环境基因组文库生产
JoVE Journal
Biology
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JoVE Journal Biology
Large Insert Environmental Genomic Library Production
DOI:

20:59 min

September 23, 2009

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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

Summary

Automatic Translation

是描述一个与环境基因组DNA经季节性缺氧峡湾的垂直深度连续分离的fosmid库建设。由此产生的克隆库回升到384孔板和存档,为下游的测序和一个自动化殖民地采摘系统的应用功能筛选。

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