{"@type": "dcat:Dataset", "accessLevel": "public", "bureauCode": ["009:25"], "contactPoint": {"@type": "vcard:Contact", "fn": "NIH", "hasEmail": "mailto:info@nih.gov"}, "description": "Background\n          We describe an alternative method to determine mRNA half-life (t1/2) based on the Real-Time RT-PCR procedure. This approach was evaluated by using the \u03b2-actin gene as a reference molecule for measuring of mRNA stability.\n        \n        \n          Results\n          Human leukemia Nalm-6 and CCRF-CEM cells were treated with various concentrations of Actinomycin D to block transcription and aliquots were removed periodically. Total RNA was isolated and quantified using the RiboGreen\u00ae fluorescent dye with the VersaFluor Fluorometer System. One \u03bcg of total RNA was reverse transcribed and used as template for the amplification of a region of the \u03b2-actin gene (231 bp). To generate the standard curve, serial ten-fold dilutions of the pBactin-231 vector containing the cDNA amplified fragment were employed, \u03b2-actin mRNAs were quantified by Real-Time RT-PCR using the SYBR\u00ae Green I fluorogenic dye and data analyzed using the iCycle iQ system software. Using this method, the \u03b2-actin mRNA exhibited a half-life of 6.6 h and 13.5 h in Nalm-6 and CCRF-CEM cells, respectively. The t1/2 value obtained for Nalm-6 is comparable to those estimated from Northern blot studies, using normal human leukocytes (5.5 h).\n        \n        \n          Conclusions\n          We have developed a rapid, sensitive, and reliable method based on Real-Time RT-PCR for measuring mRNA half-life. Our results confirm that \u03b2-actin mRNA half-life can be affected by the cellular growth rate.", "distribution": [{"@type": "dcat:Distribution", "description": "Visit the original government dataset for complete information, documentation, and data access.", "downloadURL": "https://www.ncbi.nlm.nih.gov/pmc/articles/PMC116431/", "mediaType": "text/html", "title": "Official Government Data Source"}], "identifier": "https://healthdata.gov/api/views/vd57-3u37", "issued": "2025-07-14", "keyword": ["beta-actin", "leukemia-cell-lines", "mrna-decay", "nih", "rt-pcr"], "landingPage": "https://healthdata.gov/d/vd57-3u37", "modified": "2025-09-06", "programCode": ["009:033"], "publisher": {"@type": "org:Organization", "name": "National Institutes of Health"}, "theme": ["NIH"], "title": "Real-time RT-PCR analysis of mRNA decay: half-life of Beta-actin mRNA in human leukemia CCRF-CEM and Nalm-6 cell lines"}