Improved RNA isolation from cells in tissue culture using a commercial nucleic acid extractor.
The Applied Biosystems 340A Nucleic Acid Extractor automates isolation of either DNA or RNA from tissue or cells in culture. We have found that several modifications to the manufacturer's recommended protocol greatly improve the quality of RNA that can be routinely isolated from cells in culture. These modifications include lysis of monolayer cells directly on plates, centrifuging samples after homogenization to remove precipitable RNase contaminants and purging the instrument's reagent lines with 0.1% diethyl pyrocarbonate. These simple modifications enhance both RNA quality and reproducibility of yield.
Briscoe
P R
PR
Dept. of Radiation Medicine, Vincent T. Lombardi Cancer Research Center, Georgetown University Medical Center, Washington, DC 20007.
Jorgensen
T J
TJ
D015152:Blotting, Northern; D002498:Centrifugation; D046508:Culture Techniques; D004587:Electrophoresis, Agar Gel; D019286:Endopeptidase K; D004866:Equipment Contamination; D012313:RNA / Q000302:isolation & purification*; D012697:Serine Endopeptidases
D012313:RNA; D012697:Serine Endopeptidases; D019286:Endopeptidase K