Determination of the poly(A) tail lengths of a single mRNA species in total hepatic RNA

Biotechniques. 1995 Mar;18(3):465-9.

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.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • DNA Probes / genetics
  • Electrophoresis, Polyacrylamide Gel
  • Humans
  • Liver
  • Poly A / genetics*
  • RNA, Messenger / genetics*
  • Rats
  • Ribonuclease H / metabolism
  • Sequence Analysis, RNA

Substances

  • DNA Probes
  • RNA, Messenger
  • Poly A
  • Ribonuclease H