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Streamlined Single Cell TCR Isolation and Generation of Retroviral Vectors for In Vitro and In Vivo Expression of Human TCRs
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JoVE 杂志 免疫与感染
Streamlined Single Cell TCR Isolation and Generation of Retroviral Vectors for In Vitro and In Vivo Expression of Human TCRs

Streamlined Single Cell TCR Isolation and Generation of Retroviral Vectors for In Vitro and In Vivo Expression of Human TCRs

DOI:
10.3791/55379-v

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11:21 min

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September 10, 2017

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, , , , , , , ,
1Department of Pediatrics, Section of Diabetes and Endocrinology, McNair Medical Institute,Baylor College of Medicine, Texas Children’s Hospital, 2Benaroya Research Institute at Virginia Mason, 3Center for Human Immunobiology,Baylor College of Medicine, Texas Children’s Hospital, 4Department of Pediatrics, Section of Diabetes and Endocrinology,Baylor College of Medicine, Texas Children’s Hospital

Chapters

  • 00:00Title
  • 01:42Single cell sort and reverse transcription reaction
  • 03:37Multiplex-nested PCR
  • 08:56Subcloning TCR alpha and beta chains
  • 09:56Verification of TCR expression
  • 10:46Conclusion

Summary

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The current protocol combines single cell paired human TCR alpha and beta chain sequencing with streamlined generation of retroviral vectors compatible with in vitro and in vivo TCR expression.

Tags

Keywords: Single Cell TCR IsolationTCR Alpha And Beta RepertoireTCR CloningMultiplex PCRNested PCRChimeric TCR ConstructSingle-cell PCRReverse TranscriptionT Cell Sorting

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