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T4 RNA ligase has been used to construct a series of defined oligoribonucleotides. Hexamer or pentamer blocks were synthesized first by multiple additions of mononucleotide diphosphates to trimers with T4 RNA ligase and removal of the terminal phosphate with alkaline phosphatase; inhibitors of the ligase were removed by passing the sample over a 1-ml reverse-phase octadecasilyl column. The two nucleotide blocks were then ligated to give undecamers. Yields for the individual ligations ranged from 85 to 100% for acceptors lacking uridines and at least 70% for those containing uridines. The overall yield of the undecamer relative to the starting trimers was about 10%. Each round of ligation averaged about 8 h; the time required to synthesize each undecamer was 1 to 2 weeks. Optimization of the steps to achieve this is described in detail.
T Middleton, and
W C Herlihy, and
P R Schimmel, and
H N Munro
January 1995,
Biochimie,
T Middleton, and
W C Herlihy, and
P R Schimmel, and
H N Munro
June 1991,
Bioorganicheskaia khimiia,
T Middleton, and
W C Herlihy, and
P R Schimmel, and
H N Munro
January 1977,
Nucleic acids research,
T Middleton, and
W C Herlihy, and
P R Schimmel, and
H N Munro
January 1975,
Proceedings of the National Academy of Sciences of the United States of America,
T Middleton, and
W C Herlihy, and
P R Schimmel, and
H N Munro
January 1979,
FEBS letters,
T Middleton, and
W C Herlihy, and
P R Schimmel, and
H N Munro
May 1978,
Biochemistry,
T Middleton, and
W C Herlihy, and
P R Schimmel, and
H N Munro
September 1982,
Analytical biochemistry,
T Middleton, and
W C Herlihy, and
P R Schimmel, and
H N Munro
March 1979,
Biochimica et biophysica acta,
T Middleton, and
W C Herlihy, and
P R Schimmel, and
H N Munro
October 1972,
Proceedings of the National Academy of Sciences of the United States of America,
T Middleton, and
W C Herlihy, and
P R Schimmel, and
H N Munro
May 1976,
Archives of biochemistry and biophysics,
