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Kapitel
- 00:05Titel
- 00:39Generation of Retroviral Stock
- 02:20Viral Infection of HEK293T Cells
- 04:30Genomic DNA Fragmentation
- 05:33Ligation of Linker Oligonucleotide to Fragmented Genomic DNA
- 06:58Amplification of Viral LTR-host Genomic DNA Junctions by Semi-nested PCR
- 08:02Results: Sequence Logos Depicting HIV-1 Base Preferences from Representative Experiment Libraries
- 08:52Conclusion
We describe a protocol for amplifying retroviral integration sites from the genomic DNA of infected cells, sequencing the amplified virus-host junctions, and then mapping these sequences to a reference genome. We also describe techniques to quantify the distribution of integration sites relative to various genomic annotations using BEDTools.
