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 often impossible to rapidly identify by flow cytometry the T-cell receptor variable genes in patients suffering from T-cell malignancies. This applies especially to the alpha variable genes (TRAV), since only very few anti-TcR variable alpha mAb are available. We describe a very rapid method for inverse PCR amplification of the TcR alpha chain without prior purification of the double-stranded cDNA, provide the sequences for appropriate oligonucleotides, and describe a buffer system that dramatically enhances the amplification efficiency as compared to standard conditions.
Hinz
T
T
Paul-Ehrlich-Institute, Department of Immunology, Paul-Ehrlich-Strasse 51-59, D-63225, Langen, Germany. hinth@pei.de
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D002021:Buffers; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D005784:Gene Amplification; D006801:Humans; D016399:Lymphoma, T-Cell / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics*
D002021:Buffers; D018076:DNA, Complementary; D016693:Receptors, Antigen, T-Cell, alpha-beta