Preparative in vitro mRNA synthesis using SP6 and T7 RNA polymerases.
A method for preparative in vitro mRNA synthesis is presented. The method makes it possible to generate several hundred RNA copies per molecule of linearized DNA template. The protocol is applicable to the synthesis of RNAs of differing lengths (from 200 to 3000 bases), and both SP6 and T7 RNA polymerases can be used. The transcription can be scaled up to at least 6-12 ml, yielding 4.5-18 mg of homogeneous biologically active mRNA.
Gurevich
V V
VV
Branch of M. M. Shemyakin Institute of Bioorganic Chemistry, Academy of Sciences of the USSR, Pushchino, Moscow Region.
Pokrovskaya
I D
ID
Obukhova
T A
TA
Zozulya
S A
SA
D012321:DNA-Directed RNA Polymerases / Q000494:pharmacology*; D004926:Escherichia coli / Q000201:enzymology; D005818:Genetic Engineering; D012333:RNA, Messenger / Q000138:chemical synthesis*; D014158:Transcription, Genetic
D012333:RNA, Messenger; D012321:DNA-Directed RNA Polymerases