title: Isolation of Nascent Transcripts with Click Chemistry.
abstract: Steady-state levels of cellular RNA are determined by both transcriptional rate and RNA half-life. Commonly used methods for transcriptional analysis are only capable of profiling total RNA and do not distinguish changes in synthesis and decay rates. Hence, a better understanding of the temporal dynamics of cellular response for a given condition at the transcriptional level requires techniques for the analysis of nascent transcripts. Here we describe a protocol that allows isolation of nascent transcripts with a copper-catalyzed azide-alkyne cycloaddition (CuAAC) also known as a click chemistry reaction.
authors.affiliation: Department of Molecular Biology, Massachusetts General Hospital, Boston, Massachusetts.|Department of Genetics, Harvard Medical School, Boston, Massachusetts.
authors.forename: Ozlem
authors.initials: O
authors.lastname: Yildirim
mesh_terms: D057930:Click Chemistry / Q000379:methods*; D003584:Cytological Techniques / Q000379:methods*; D020869:Gene Expression Profiling / Q000379:methods*; D012322:RNA Precursors / Q000302:isolation & purification*; D012333:RNA, Messenger / Q000302:isolation & purification*
chemical_list: D012322:RNA Precursors; D012333:RNA, Messenger
keywords: click chemistry; metabolic labeling; nascent RNA