Applications of magnetic beads with covalently attached oligonucleotides in hybridization: isolation and detection of specific measles virus mRNA from a crude cell lysate

Anal Biochem. 1990 Aug 15;189(1):40-50. doi: 10.1016/0003-2697(90)90041-7.

Abstract

A novel, rapid, one-step isolation procedure utilizing oligonucleotides end-attached to magnetic beads (Dynabeads) has been developed. The beads and their captured target nucleic acids were separated after 2 h from the hybridization solution using an external magnet. This procedure was used to isolate measles virus hemagglutinin (H) mRNA from infected cells dissolved in 5 M guanidine thiocyanate. The yield of the specific, undegraded H-mRNA was found to be near the estimated total amount of H-mRNA present in the cells. The magnetic beads were also used for detection of measles virus H-mRNA in a crude cell lysate by sandwich hybridization. The experimental detection limit was 630 amol H-mRNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Magnetics
  • Molecular Sequence Data
  • Nucleic Acid Hybridization*
  • Nucleic Acids / isolation & purification
  • Nucleic Acids / metabolism
  • Oligonucleotides / chemistry
  • Oligonucleotides / genetics
  • RNA, Messenger / isolation & purification
  • RNA, Viral / isolation & purification

Substances

  • Nucleic Acids
  • Oligonucleotides
  • RNA, Messenger
  • RNA, Viral