A simple and efficient method for the isolation of total RNA from the fission yeast Schizosaccharomyces pombe.
A simple and efficient procedure was described for the isolation of total RNA from the fission yeast Schizosaccharomyces pombe. The present study demonstrated that the quality and the quantity of S. pombe RNA were increased by substituting phenol/chloroform mixture for phenol as a deproteinizing agent in the first vortexing step and using an ice bath instead of a dry ice-ethanol bath in the freezing step. Additionally, this protocol had the advantage of extracting total RNA without any degradation of S. pombe cells. Furthermore, the high amounts and quality of RNA extracted by this modified procedure enabled us to perform some experiments such as Northern blot, S1 mapping, primer extension, and reverse transcriptase reaction-polymerase chain reaction (RT-PCR) without further RNA purification. We suggest that this procedure is very useful to analyse primary structures and steady-state levels of RNA from S. pombe.
Jang
Y K
YK
Department of Molecular Biology, Seoul National University, Republic of Korea.
Jin
Y H
YH
Kim
M J
MJ
Seong
R H
RH
Hong
S H
SH
Park
S D
SD
D001483:Base Sequence; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012331:RNA, Fungal / Q000302:isolation & purification*; D012568:Schizosaccharomyces / Q000737:chemistry / Q000235:genetics*
D012331:RNA, Fungal