Wt. Stump et Kb. Hall, SP6 RNA-POLYMERASE EFFICIENTLY SYNTHESIZES RNA FROM SHORT DOUBLE-STRANDED DNA TEMPLATES, Nucleic acids research, 21(23), 1993, pp. 5480-5484
SP6 DNA-dependent RNA polymerase, like T7 RNA polymerase, can be used
to synthesize RNA sequences from short DNA templates which contain the
1 8 base pair promoter region. Use of SP6 polymerase extends the rang
e of possible 5' sequences of RNA products, since the preferred SP6 st
art site (of the RNA product) is 5'GAAGA, while T7 polymerase prefers
5'GGGAG. The SP6 start site can be advantageous in large-scale synthes
es where high concentrations of RNA can lead to aggregation. Using the
limited number of DNA templates described here, there appears to be a
significant difference between the two enzymes: SP6 polymerase requir
es a complete duplex DNA substrate for efficient synthesis, unlike the
T7 enzyme which works efficiently when only the 18 base promoter regi
on is double-stranded. SP6 polymerase consistently produces higher yie
lds of RNA than does T7 polymerase, and the reactions can be easily sc
aled up to produce milligram quantities of RNA.