Application of reverse transcription PCR and capillary electrophoresis in the analysis of T-cell receptor beta variable repertoire

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Authors

TESAŘOVÁ Lenka PETERKOVÁ Martina STEJSKAL Stanislav POTĚŠILOVÁ Michaela KRONTORÁD KOUTNÁ Irena

Year of publication 2010
Type Article in Proceedings
Conference Advances in Molecular and Cancer Biology, 2010
MU Faculty or unit

Faculty of Informatics

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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