An improved RT-PCR protocol for the quantitation of human retinoic acid receptor RNA.
A quantitative reverse transcriptase polymerase chain reaction (RT-PCR) system has been developed to calculate the level of expression of human retinoic acid receptors (hRAR) alpha, beta, and gamma. Starting from a single cDNA preparation, the system allows the measurement of the number of molecules of each mRNA receptor. This is made possible by a synthetic internal standard mRNA which is added in known concentrations at the beginning of the reaction. The system is tested in a rhabdomyosarcoma cell line (A-673) where we have measured the upregulation of beta and gamma receptor mRNAs following treatment with retinoic acid.
Ferrari
N
N
Laboratory of Molecular Biology, Istituto Nazionale per la Ricerca sul Cancro, Genova, Italy.
Pfeffer
U
U
Tosetti
F
F
Brigati
C
C
Vidali
G
G
D001483:Base Sequence; D004273:DNA, Neoplasm / Q000032:analysis / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012313:RNA / Q000032:analysis* / Q000235:genetics; D018168:Receptors, Retinoic Acid / Q000032:analysis / Q000235:genetics*; D015203:Reproducibility of Results; D012208:Rhabdomyosarcoma / Q000737:chemistry / Q000473:pathology / Q000648:ultrastructure; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured; D015854:Up-Regulation
D004273:DNA, Neoplasm; D018168:Receptors, Retinoic Acid; D012313:RNA