TY - JOUR
T1 - Preparative-scale purification of RNA using an efficient method which combines gel electrophoresis and column chromatography
AU - Cunningham, Lynette
AU - Kittikamron, Kirk
AU - Lu, Yi
N1 - Funding Information:
We thank J. J. Dunn and A. H. Rosenberg for providing the plasmid pAR1219 containing the T7 RNA polymerase gene, Gretchen Peterson for purification of the T7 RNA polymerase, Pascale Legault and Arthur Pardi for providing the RNA transcription protocol, Daniel Celander, Olke Uhlenbeck and Harry Noller for helpful suggestions, and Richard Gumport for use of gel digitalization equipment and critical reading of the manuscript. This research was supported by the NIH FIRST Award (GM53706), the Beckman Young Investigator Award, and the Donors of the Petroleum Research Fund, administered by the American Chemical Society.
PY - 1996
Y1 - 1996
N2 - Here we describe a reliable method for purifying large amounts of RNA of any sequence and length with comparable efficiency and resolution of gel electrophoresis and with capacity approaching that of column chromatography. The RNA mixture of interest is separated on a cylindrical denaturing polyacrylamide gel, eluted by a peristaltic pump, detected by a UV-vis detector, and collected by a fraction collector. Using this method, we were able to separate one third of a 100 ml in vitro transcribed 34mer hammerhead ribozyme (~6.2 mg) in a single run. The entire 100 ml transcribed RNA (~18.5 mg) was separated after consecutive runs using one single gel preparation.
AB - Here we describe a reliable method for purifying large amounts of RNA of any sequence and length with comparable efficiency and resolution of gel electrophoresis and with capacity approaching that of column chromatography. The RNA mixture of interest is separated on a cylindrical denaturing polyacrylamide gel, eluted by a peristaltic pump, detected by a UV-vis detector, and collected by a fraction collector. Using this method, we were able to separate one third of a 100 ml in vitro transcribed 34mer hammerhead ribozyme (~6.2 mg) in a single run. The entire 100 ml transcribed RNA (~18.5 mg) was separated after consecutive runs using one single gel preparation.
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U2 - 10.1093/nar/24.18.3647
DO - 10.1093/nar/24.18.3647
M3 - Article
C2 - 8836197
AN - SCOPUS:0029844437
SN - 0305-1048
VL - 24
SP - 3647
EP - 3648
JO - Nucleic acids research
JF - Nucleic acids research
IS - 18
ER -