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T and B Cell Receptor Immune Repertoire Analysis using Next-generation Sequencing
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Immunology and Infection
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JoVE Journal Immunology and Infection
T and B Cell Receptor Immune Repertoire Analysis using Next-generation Sequencing

T and B Cell Receptor Immune Repertoire Analysis using Next-generation Sequencing

DOI:
10.3791/61792-v

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08:59 min

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January 12, 2021

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, , , ,
1Pediatric Gastroenterology Unit, Edmond and Lily Safra Children’s Hospital,Sheba Medical Center, 2Sackler Faculty of Medicine,Tel Aviv University, 3Cancer Research Center,Sheba Medical Center, 4Molecular Hematology Laboratory,Sheba Medical Center

Chapters

  • 00:05Introduction
  • 00:48DNA Isolation and Library Preparation
  • 04:21Amplicon Purification and Quantification
  • 05:57Next-generation Sequencing
  • 06:57Results: TCRβ and IGH Immune Repertoire Data Analysis
  • 08:27Conclusion

Summary

Automatic Translation

Automatic Translation

The current protocol describes a method for DNA isolation from blood samples and intestinal biopsies, generation of TCRβ and IGH PCR libraries for next-generation sequencing, performance of a NGS run and basic data analysis.

Tags

T Cell ReceptorB Cell ReceptorNext-generation SequencingLymphocyte DiversityBiomarkersDNA IsolationCell LysisProteinase KProtein PrecipitationDNA Extraction

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