Regular ArticleTranscriptional Pulsing Approaches for Analysis of mRNA Turnover in Mammalian Cells☆
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Renal Ammonium Ion Production and Excretion
2013, Seldin and Geibisch's The KidneyRenal Ammonium Ion Production and Excretion
2012, Seldin and Giebisch's The Kidney: Physiology and PathophysiologyChapter 24 Assays for Determining Poly(A) Tail Length and the Polarity of mRNA Decay in Mammalian Cells
2008, Methods in EnzymologyChapter 17 Messenger RNA Half-Life Measurements in Mammalian Cells
2008, Methods in EnzymologyCitation Excerpt :After transient transfection of the Tet‐off promoter–driven reporter plasmid, the transfected cells should be kept in culture medium containing the identified concentration of tetracycline until transcription induction (transcriptional pulsing). It is striking that all three different mammalian stable lines we established, including mouse NIH3T3 B2A2, human K562 III‐2, and human BEAS‐2B‐19, show the same transcriptional pulsing kinetics in Northern blot analysis (Chen et al., 2007; Loflin et al., 1999b; Xu et al., 1998). A population of mRNA homogenous in size appears in the cytoplasm after removal of tetracycline for 100 to 110 min, making it possible to study deadenylation and decay kinetics and precursor and product relationships during mRNA decay.
Chapter 22 Sensitive Detection of mRNA Decay Products by Use of Reverse-Ligation-Mediated PCR (RL-PCR)
2008, Methods in Enzymology
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Lefkovits, I.
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