Biopolym. Cell. 1990; 6(3):50-55.
http://dx.doi.org/10.7124/bc.00026B
Structure and Function of Biopolymers
Preparative synthesis of ribooligonucleotides with the preset sequence by transcription of deoxyribonucleotides
1Denisova L. Ya., 2Zagrebelny S. N., 1Pustoshilova N. M., 1Venyaminova A. G., 2Corn V. V., 2Zaritova V. F., 2Repkova M. N.
  1. Research and Technology Institute of Biologically Active Substances
    Berdsk, Novosibirsk Region, USSR
  2. Institute of Bioorganic Chemistry, Siberian Branch of the Academy of Sciences of the USSR
    Novosibirsk, USSR

Abstract

The transcription of complementary deoxyribooligonucleotides was performed to obtain AGGGGAUUGAAAAUC (tRNAphe anticodon loop fragment) and AUGAGGAAUACCCAUG (the MS2 phase RNA fragment) in preparative amounts. The nucleotide incorporation into the transcription product was shown to achieve 70 % of the template nucleotide level. The procedure was developed for the isolation of the transcript from the reaction mixture. The isolation steps involve geparine-agarose chromatography, Lichroprep RP 18 one and the electrophoresis in 20 % polyacrylamide gel under denaturation conditions followed by electroelution of the desired product from gel. Being an exact copy of deoxyribotemplate the ribooligonucleotide gave the yield of 1.0–1.5 U260 on 5.0 U260 of the template.
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