Additional TCRV beta primers and minor method modifications improve detection of clonal T-cell populations by RT-PCR
| Autor: | D Howe, C Lynas |
|---|---|
| Rok vydání: | 1997 |
| Předmět: |
DNA
Complementary Hot Temperature Receptors Antigen T-Cell alpha-beta T cell Population Immunoglobulin Variable Region Biology Polymerase Chain Reaction Pathology and Forensic Medicine law.invention T-Lymphocyte Subsets law Complementary DNA medicine education Polymerase chain reaction Genetics education.field_of_study Molecular biology Reverse transcriptase Clone Cells Real-time polymerase chain reaction medicine.anatomical_structure Polyclonal antibodies biology.protein Electrophoresis Polyacrylamide Gel Primer (molecular biology) Research Article |
| Zdroj: | Molecular Pathology. 50:53-55 |
| ISSN: | 1366-8714 |
| DOI: | 10.1136/mp.50.1.53 |
| Popis: | The TCRV beta RT-PCR method for detection of clonal populations of T cells which we described previously could not detect clones that used certain variable (V) beta region families. V beta 2, 4, 8.3, and 18 had insufficient homology with the original consensus V region primer. Two new primers have been designed which work well and are able to amplify from V beta families previously undetectable by this RT-PCR. In addition, minor alterations to the cDNA synthesis and gel analysis of the PCR products make the results even easier to interpret. All the Diversity/Joining (D/J) region primer combinations except for D2/J2 have been omitted, and terminating the reverse transcription by heating prior to PCR greatly improves amplification with these primers. Use of 8% and/or 10% polyacrylamide gels increases clarity. Inclusion of the modifications described will reduce false reporting of patients as having a polyclonal T-cell population. |
| Databáze: | OpenAIRE |
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