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Nucleic Acids Research, 1985, Vol. 13, No. 21 7663-7672
© 1985

Articles

Selective protection of in vitro synthesized cDNA against nucleases by incorporation of phosphorothioate-analogues

Georg Schreiber, Edith M. Koch and Wolfgang J. Neubert

Max-Planck-Institut für Biochemie, Abteilung für Virusforschung 8033 Martinsried bei München, FRG

Received August 5, 1985. Revised September 26, 1985. Accepted October 15, 1985.

The conditions for the stepwise synthesis of single- (ss) and double-stranded (ds) cDNA using thio-analogues instead of dNTPs are described in this paper. RNA of paramyxovirus Sendai (strain 6/94) aervea as template in these experiments. The increased resistance of this {propto}S-modified cDNA against several nucleases, like Sl-Nuclease, DNase I, Exonuclease III, snake venom Phosphodiesterase (PDE) and the combination of DNase I and PDE is demonstrated.


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