title: Determination of the poly(A) tail lengths of a single mRNA species in total hepatic RNA.
abstract: Poly(A) tail length is important in the stability and translation of mRNA. We describe procedures for the rapid and reproducible analysis of poly(A) tail length of a single mRNA species contained in a sample of total hepatic RNA. A short 3' fragment of a specific mRNA is prepared by RNase H digestion of the targeted mRNA region annealed to a short DNA oligonucleotide. The length of the poly(A) tail of the 3' fragment is then determined by running the sample on a polyacrylamide gel, by electrophoretic transfer, by probing with a radiolabeled cDNA and by comparing the size of the detected region with a specific RNA ladder or a DNA ladder.
authors.affiliation: Institute of Chemical Toxicology, Wayne State University, Detroit, MI 48201, USA.
authors.forename: R C
authors.initials: RC
authors.lastname: Zangar
authors.forename: M
authors.initials: M
authors.lastname: Hernandez
authors.forename: T A
authors.initials: TA
authors.lastname: Kocarek
authors.forename: R F
authors.initials: RF
authors.lastname: Novak
mesh_terms: D000818:Animals; D015342:DNA Probes / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008099:Liver; D011061:Poly A / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics*; D051381:Rats; D016914:Ribonuclease H / Q000378:metabolism; D017423:Sequence Analysis, RNA
chemical_list: D015342:DNA Probes; D012333:RNA, Messenger; D011061:Poly A; D016914:Ribonuclease H