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Amplification of Near Full-length HIV-1 Proviruses for Next-Generation Sequencing
JoVE Journal
Genetics
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JoVE Journal Genetics
Amplification of Near Full-length HIV-1 Proviruses for Next-Generation Sequencing

Amplification of Near Full-length HIV-1 Proviruses for Next-Generation Sequencing

DOI:
10.3791/58016-v

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10:18 min

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October 16, 2018

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, , ,
1Centre for Virus Research, The Westmead Institute for Medical Research,University of Sydney

Chapters

  • 00:04Title
  • 00:40Lysis of HIV-1-infected Cells and Amplification of Single HIV-1 DNA Proviruses via Nested PCR
  • 04:23DNA Purification and Quantification
  • 08:12Results: Analysis of Amplified Near Full-length HIV-1 Proviruses
  • 09:21Conclusion

Summary

Automatic Translation

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Full-length individual proviral sequencing (FLIPS) provides an efficient and high-throughput method for the amplification and sequencing of single, near full-length (intact and defective) HIV-1 proviruses and allows for determination of their potential replication-competency. FLIPS overcomes limitations of previous assays designed to sequence the latent HIV-1 reservoir.

Tags

Keywords: HIV-1 ProvirusesNext-generation SequencingSingle-cellAmplificationPCRGenetic CompositionReplication CompetentCell TypesLysis BufferSerial DilutionPositive ControlThermal Cycler

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