Abstract
This protocol describes the quantification of all expressed T-cell antigen receptor (TCR) gene products within sorted (by flow cytometry) EBV and CMV-specific memory CD8+ T-cell populations using a template-switch anchored reverse transcription polymerase chain reaction (RT-PCR).
Materials and Reagents
Equipment
Procedure
Representative data
Example of a TCRβ-PCR: Figure 1. A duplicate TCRβ-PCR was performed on 3 different donors. A 1 kb Plus DNA ladder was used. The correct PCR product is ±550 bp long. Note the presence of PCR products other than the desired one, which is a common side effect of SMART cDNA synthesis.
Notes
Recipes
Acknowledgments
This protocol was adapted from the previously published studies (Douek et al., 2002; Quigley et al., 2011).
References
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