Synthesis of digoxigenin-labeled cDNA as a test for mRNA integrity.
The nucleotide analogue digoxigenin-11-dUTP is widely used as a nonradioactive marker in a broad range of techniques including dot blots, Southern and Northern blots, colony and plaque screenings and in situ hybridizations. In this report, we describe the incorporation of this molecule into a cDNA synthesized by reverse transcriptase as a novel application for digoxigenin. This reaction can be performed as a useful control to check the integrity of a bulk mRNA preparation in the construction of a cDNA library, since the ability of mRNA to direct the synthesis of long molecules of first-strand cDNA is a sign of its integrity.
D004247:DNA / Q000096:biosynthesis* / Q000235:genetics; D015342:DNA Probes; D003856:Deoxyuracil Nucleotides; D004076:Digoxigenin / Q000031:analogs & derivatives*; D005069:Evaluation Studies as Topic; D005821:Genetic Techniques; D015336:Molecular Probe Techniques; D012333:RNA, Messenger / Q000235:genetics*; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism
D015342:DNA Probes; D003856:Deoxyuracil Nucleotides; D012333:RNA, Messenger; C070202:digoxigenin-11-deoxyuridine triphosphate; D004247:DNA; D012194:RNA-Directed DNA Polymerase; D004076:Digoxigenin