title: A modified DNA isolation protocol for obtaining pure RT-PCR grade RNA.
abstract: 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.
authors.affiliation: Institut für Molekulare Mikrobiologie und Biotechnologie, Westfälische Wilhelms-Universität Münster, Corrensstr. 3, 48149, Munster, Germany. roland.klassen@uni-muenster.de
authors.forename: Roland
authors.initials: R
authors.lastname: Klassen
authors.forename: Julia
authors.initials: J
authors.lastname: Fricke
authors.forename: Annika
authors.initials: A
authors.lastname: Pfeiffer
authors.forename: Friedhelm
authors.initials: F
authors.lastname: Meinhardt
mesh_terms: 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
chemical_list: 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