A fast and simple method to determine the clonal relationship among human T-cell lymphocytes.
For the analysis of mutation in human T-cell lymphocytes, it is crucial to determine the clonal relationship between isolated mutants, particularly if they harbour identical mutations. Here we report an efficient method to determine the clonal relationships between these cells. The method is based on the analysis of restriction fragment length polymorphisms of a polymerase chain reaction amplified, rearranged T-cell receptor gamma-gene. As few as 600 cells are sufficient, regular agarose gels can be used for the separation of the restriction fragments, no radioactive label is required, and results can be obtained in 2 days.
de Boer
J G
JG
Biology Department, University of Victoria, BC, Canada.
Curry
J D
JD
Glickman
B W
BW
D001483:Base Sequence; D002999:Clone Cells; D004247:DNA / Q000032:analysis; D004252:DNA Mutational Analysis / Q000379:methods; D004587:Electrophoresis, Agar Gel; D015334:Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000235:genetics*; D015203:Reproducibility of Results; D013601:T-Lymphocytes / Q000276:immunology*
D016692:Receptors, Antigen, T-Cell, gamma-delta; D004247:DNA