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Identifying DNA Mutations in Purified Hematopoietic Stem/Progenitor Cells
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Immunology and Infection
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JoVE Journal Immunology and Infection
Identifying DNA Mutations in Purified Hematopoietic Stem/Progenitor Cells

Identifying DNA Mutations in Purified Hematopoietic Stem/Progenitor Cells

DOI:
10.3791/50752-v

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11:06 min

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February 24, 2014

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, , , , , , ,
1Greehey Children’s Cancer Research Institute,UT Health Science Center at San Antonio, 2Department of Cellular and Structural Biology,UT Health Science Center at San Antonio, 3Department of Pathology,UT Health Science Center at San Antonio, 4Department of Microbiology,UT Health Science Center at San Antonio, 5Cancer Therapy and Research Center,UT Health Science Center at San Antonio

Chapters

  • 00:05Title
  • 01:45Isolation of Genomic DNA
  • 04:10Packaging of Genomic DNA
  • 05:21Plating Packaged Genomic DNA
  • 07:07Determination of a Putative Mutant Frequency
  • 08:37Results: The in vivo Mutagenesis Assay
  • 10:35Conclusion

Summary

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Here we describe an in vivo mutagenesis assay for small numbers of purified hematopoietic cells using the LacI transgenic mouse model. The LacI gene can be isolated to determine the frequency, location, and type of DNA mutants spontaneously arisen or after exposure to genotoxins.

Tags

Keywords: DNA MutationsHematopoietic Stem CellsHematopoietic Progenitor CellsGenomic InstabilityLacI Transgenic MiceIn Vivo Mutagenesis AssayLin-IL7R-Sca-1+cKit++ (LSK) CellsDNA SequencingMutation Frequency

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