Identification of the T-cell receptor alpha variable (TRAV) gene(s) in T-cell malignancies

Due to the lack of a complete range of monoclonal antibodies (mAb) it is of ten impossible to rapidly identify by flow cytometry the T-cell receptor va riable genes in patients suffering from T-cell malignancies. This applies e specially to the alpha variable genes (TRAV), since only very few anti-TcR variable alpha mAb are available. We describe a very rapid method for inver se PCR amplification of the TcR alpha chain without prior purification of t he double-stranded cDNA, provide the sequences for appropriate oligonucleot ides, and describe a buffer system that dramatically enhances the amplifica tion efficiency as compared to standard conditions. (C) 2000 Elsevier Scien ce B.V. All rights reserved.