Synthesis of digoxigenin-labeled cDNA as a test for mRNA integrity.

Biotechniques

Istituto Ricerca F. Angelini S.p.A. P.le della Stazione s.n.c., Pomezia, Rome, Italy.

Published: December 1992

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.

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