-transparent.png){kind=logo}
A method is described to synthesize small RNAs of defined length and sequence using T7 RNA polymerase and templates of synthetic DNA which contain the T7 promoter. Partially single stranded templates which are base paired only in the -17 to +1 promoter region are just as active in transcription as linear plasmid DNA. Runoff transcripts initiate at a unique, predictable position, but may have one nucleotide more or less on the 3' terminus. In addition to the full length products, the reactions also yield a large amount of smaller oligoribonucleotides in the range from 2 to 6 nucleotides which appear to be the result of abortive initiation events. Variants in the +1 to +6 region of the promoter are transcribed with reduced efficiency but increase the variety of RNAs which can be made. Transcription reaction conditions have been optimized to allow the synthesis of milligram amounts of virtually any RNA from 12 to 35 nucleotides in length.
J F Milligan, and
D R Groebe, and
G W Witherell, and
O C Uhlenbeck
February 1989,
Bioorganicheskaia khimiia,
J F Milligan, and
D R Groebe, and
G W Witherell, and
O C Uhlenbeck
November 2013,
Cold Spring Harbor protocols,
J F Milligan, and
D R Groebe, and
G W Witherell, and
O C Uhlenbeck
January 2023,
Methods in enzymology,
J F Milligan, and
D R Groebe, and
G W Witherell, and
O C Uhlenbeck
October 1978,
Indian journal of biochemistry & biophysics,
J F Milligan, and
D R Groebe, and
G W Witherell, and
O C Uhlenbeck
August 1998,
Nucleic acids research,
J F Milligan, and
D R Groebe, and
G W Witherell, and
O C Uhlenbeck
March 2022,
Biotechnology journal,
J F Milligan, and
D R Groebe, and
G W Witherell, and
O C Uhlenbeck
June 1991,
Carcinogenesis,
J F Milligan, and
D R Groebe, and
G W Witherell, and
O C Uhlenbeck
March 1976,
Nature,
J F Milligan, and
D R Groebe, and
G W Witherell, and
O C Uhlenbeck
July 1973,
Biochimica et biophysica acta,
J F Milligan, and
D R Groebe, and
G W Witherell, and
O C Uhlenbeck
January 1989,
Methods in enzymology,
