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Bidirectional Retroviral Integration Site PCR Methodology and Quantitative Data Analysis Workflow
JoVE Journal
Genetics
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JoVE Journal Genetics
Bidirectional Retroviral Integration Site PCR Methodology and Quantitative Data Analysis Workflow
DOI:

12:53 min

June 14, 2017

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Chapters

  • 00:05Title
  • 01:11LTR-specific Biotin-primer Extension
  • 02:53Rsal and CviQI Digestion and Blunt-ending
  • 04:12Straptavidin Bead Binding
  • 05:31Linker Ligation
  • 06:39Pre-amplification of Both Left and Right Junction DNA
  • 08:15Left- and Right-junction-specific Amplifications
  • 10:23Results: The IS Bidirectional Assay Improves Detection Rates and Sequence Quantification Accuracy
  • 12:09Conclusion

Summary

Automatic Translation

This manuscript describes the experimental procedure and software analysis for a bidirectional integration site assay that can simultaneously analyze upstream and downstream vector-host junction DNA. Bidirectional PCR products can be used for any downstream sequencing platform. The resulting data are useful for a high-throughput, quantitative comparison of integrated DNA targets.

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