Sampling Saccharomyces cerevisiae cells by rapid filtration improves the yield of mRNAs.
To optimize the recovery of mRNAs extracted from yeast, different methods for sampling the yeast cells have been compared. For Saccharomyces cerevisiae strains growing on gluconeogenic carbon sources (derepressed cells) rapid filtration allowed much higher yields (3-10 fold) than centrifugation at room temperature or at 4 degrees C. Recovery of total RNA was similar with the different procedures. For S. cerevisiae growing on glucose, filtration caused a 2-4 fold improvement on the mRNA yields obtained from cells sampled by centrifugation. It was also observed that, when derepressed cells of S. cerevisiae W303-1A were collected by filtration and flash-frozen, part of the 25S and 18S rRNAs (up to 50%) was recovered in an unprocessed 32S or 33S form.
Belinchón
Mónica M
MM
Instituto de Investigaciones Biomédicas Alberto Sols, CSIC-UAM, Arturo Duperier 4, E-28029 Madrid, Spain.
Flores
Carmen-Lisset
CL
Gancedo
Juana M
JM
D001483:Base Sequence; D004271:DNA, Fungal / Q000235:genetics; D009172:Mycology / Q000379:methods; D012331:RNA, Fungal / Q000235:genetics / Q000302:isolation & purification*; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification*; D012335:RNA, Ribosomal / Q000235:genetics / Q000302:isolation & purification; D012337:RNA, Ribosomal, 18S / Q000235:genetics / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae / Q000737:chemistry* / Q000235:genetics / Q000254:growth & development; D013045:Species Specificity; D025062:Yarrowia / Q000737:chemistry / Q000235:genetics
D004271:DNA, Fungal; D012331:RNA, Fungal; D012333:RNA, Messenger; D012335:RNA, Ribosomal; D012337:RNA, Ribosomal, 18S; C065039:RNA, ribosomal, 25S