A modified DNA isolation protocol for obtaining pure RT-PCR grade RNA.
We provide a simple but very efficient method for RNA preparation from Saccharomyces cerevisiae based on a standard chromosomal DNA isolation protocol. The method yields DNA-free total RNA, including mRNA, rRNA, and tRNA but can easily be adjusted to considerably enrich low molecular weight RNAs, such as tRNAs and the small rRNA species (5S and 5.8S). The procedure was proven and validated by verification of cDNAs belonging to four different genes, two of which encoding polypeptides and two tRNA genes. Besides its simplicity, the method is further advantageous in terms of safety (omitting hazardous phenol) and cost efficiency.
Klassen
Roland
R
Institut für Molekulare Mikrobiologie und Biotechnologie, Westfälische Wilhelms-Universität Münster, Corrensstr. 3, 48149, Munster, Germany. roland.klassen@uni-muenster.de
Fricke
Julia
J
Pfeiffer
Annika
A
Meinhardt
Friedhelm
F
D004271:DNA, Fungal / Q000302:isolation & purification*; D003850:Deoxyribonuclease I / Q000378:metabolism; D012331:RNA, Fungal / Q000302:isolation & purification*; D012333:RNA, Messenger / Q000302:isolation & purification; D012335:RNA, Ribosomal / Q000302:isolation & purification; D012340:RNA, Ribosomal, 5.8S / Q000302:isolation & purification; D012341:RNA, Ribosomal, 5S / Q000302:isolation & purification; D012343:RNA, Transfer / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000378:metabolism
D004271:DNA, Fungal; D012331:RNA, Fungal; D012333:RNA, Messenger; D012335:RNA, Ribosomal; D012340:RNA, Ribosomal, 5.8S; D012341:RNA, Ribosomal, 5S; D012343:RNA, Transfer; D003850:Deoxyribonuclease I