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Preparation of Viral DNA from Nucleocapsids
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Immunology and Infection
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JoVE Journal Immunology and Infection
Preparation of Viral DNA from Nucleocapsids

Preparation of Viral DNA from Nucleocapsids

DOI:
10.3791/3151-v

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12:45 min

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August 16, 2011

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, ,
1Department of Molecular Biology,Princeton University

Chapters

  • 00:05Title
  • 01:34Day 1: Viral Infection and Preparation of Buffers
  • 02:56Day 2, Phase 1: Cell Lysis and Ultracentrifugation
  • 04:01Day 2, Phase 2: Ultracentrifugation
  • 07:35“Ghost” DNA Precipitation
  • 09:56Representative Results
  • 11:51Conclusion

Summary

Automatic Translation

Automatic Translation

We describe the process of isolating high purity herpesvirus nucleocapsid DNA from infected cells. The final DNA captured from solution is of high concentration and purity, making it ideally suited for high-throughput sequencing, high fidelity PCR reactions, and transfections to produce new viral recombinants.

Tags

Viral DNANucleocapsidsHost Cell ContaminantsDownstream ApplicationsViral Genome SequencingHost DNA RemovalViral Recombinant StrainsPurified PlasmidLinear Viral DNAExtractionsDensity-based CentrifugationPurified Viral CapsidsCellular ProteinsOD260/280 RatioYields Of Viral DNALong-term StorageHigh-throughput SequencingHigh Fidelity PCR ReactionsTransfections

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