Magnetic DNA hybridization properties of oligonucleotide probes attached to superparamagnetic beads and their use in the isolation of poly(A) mRNA from eukaryotic cells.
Poly(A) messenger RNA is generally purified from total RNA using oligo(dT) cellulose affinity chromatography or centrifugation through spin columns. We present a new method for rapid purification of poly(A) mRNA using oligo(dT) probes attached to superparamagnetic beads. By magnetic separation, washing, and elution, pure mRNA is obtained from living cells within 10 minutes. This procedure works for crude RNA preparations or cell lysates that would otherwise clog standard oligo(dT) cellulose column systems. The present method reduces the risk of degradation, is highly efficient, and can easily be scaled up or down.
Hornes
E
E
Apothekernes Laboratorium A.S., Oslo, Norway.
Korsnes
L
L
D000818:Animals; D001426:Bacterial Proteins; D001483:Base Sequence; D001710:Biotin; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D003593:Cytoplasm / Q000737:chemistry; D005057:Eukaryotic Cells / Q000737:chemistry*; D006868:Hydrolysis; D007700:Kinetics; D008280:Magnetics; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D011061:Poly A / Q000235:genetics*; D012333:RNA, Messenger / Q000302:isolation & purification*; D019809:Streptavidin
D001426:Bacterial Proteins; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D011061:Poly A; D001710:Biotin; D019809:Streptavidin