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Nucleic Acids Research, 1977, Vol. 4, No. 2 445-455
© 1977

Articles

On the mechanism of oligonucleotide-primed RNA synthesis. II. Synthesis of specific primer-initiated RNA copies suitable for DNA sequence analysis

C.F. Van Kreijl, R.H.J. Beelen and P. Borst

Section for Medical Enzymology and Molecular Biology, Laboratory of Biochemistry, University of Amsterdam Eerste Constantijn Huygensstraat 20, Amsterdam, The Netherlands

Received December 21, 1976. The effect of temperature and primer concentration on oligonucleotide-primed transcription has been studied using the separated strands of a well-defined natural DNA as template. Results were similar to those obtained in the homopolymer-directed model systems. At high temperature and excess primer concentration mainly primer-initiated RNA copies are synthesized. Omission of one ribonucleoside triphosphate also makes the termination specific. The unique RNA fragments thus obtained have been used to determine the perfectly-repeated sequence of 68 base pairs in this DNA


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