Extraction of nucleic acids from yeast cells and plant tissues using ethanol as medium for sample preservation and cell disruption.
Here we report that dehydrated ethanol is an excellent medium for both in situ preservation of nucleic acids and cell disruption of plant and yeast cells. Cell disruption was strongly facilitated by prior dehydration of the ethanol using dehydrated zeolite. Following removal of ethanol, nucleic acids were extracted from the homogenate pellet using denaturing buffers. The method provided DNA and RNA of high yield and integrity. Whereas cell wall disruption was essential for extraction of DNA and large RNA molecules, smaller molecules such as tRNAs could be selectively extracted from undisrupted, ethanol-treated yeast cells. Our results demonstrate the utility of absolute ethanol for sample fixation, cell membrane and cell wall disruption, as well as preservation of nucleic acids during sample storage.
Linke
Bettina
B
Department of Biology, Cell Biology, Humboldt University, Berlin, Germany.
Schröder
Kersten
K
Arter
Juliane
J
Gasperazzo
Tatiana
T
Woehlecke
Holger
H
Ehwald
Rudolf
R
D017360:Arabidopsis / Q000737:chemistry*; D004271:DNA, Fungal / Q000302:isolation & purification*; D018744:DNA, Plant / Q000302:isolation & purification*; D000431:Ethanol; D008967:Molecular Biology / Q000379:methods; D011309:Preservation, Biological; D012331:RNA, Fungal / Q000302:isolation & purification*; D018749:RNA, Plant / Q000302:isolation & purification*; D012441:Saccharomyces cerevisiae / Q000737:chemistry*
D004271:DNA, Fungal; D018744:DNA, Plant; D012331:RNA, Fungal; D018749:RNA, Plant; D000431:Ethanol