Application of reverse transcription PCR and capillary electrophoresis in the analysis of T-cell receptor beta variable repertoire
| Authors | |
|---|---|
| Year of publication | 2010 |
| Type | Article in Proceedings |
| Conference | Advances in Molecular and Cancer Biology, 2010 |
| MU Faculty or unit | |
| Citation | |
| Field | Genetics and molecular biology |
| Keywords | Reverse transcription polymerase chain reaction; Capillary electrophoresis; T-cell receptor; Beta variable families |
| Description | Monitoring the T-cell population diversity has demonstrated to be beneficial when evaluating an individuals immune status. BV families are most often analysed to examine this diversity. The aim of our study was to establish a method for specific determination of relative BV family expression and to evaluate its applicability in terms of multiple myeloma (MM). Peripheral blood mononuclear cells from healthy donors and MM patients were analysed by RT-PCR. PCR product amounts were determined with an Agilent 2100 Bioanalyzer utilizing capillary electrophoresis and converted to the relative BV family expression. This report provides reverse transcription PCR with primers allowing for the analysis of the entire BV family repertoire. Further it describes the utilization of capillary electrophoresis to evaluate the relative BV family expression after their amplification. The methodology is fast and simple to operate and is therefore suitable as a first screening of the T-cell population composition. |
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