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A modified DNA isolation protocol for obtaining pure RT-PCR grade RNA
- Klassen, Roland, Fricke, Julia, Pfeiffer, Annika, Meinhardt, Friedhelm
- Biotechnology letters 2008 v.30 no.6 pp. 1041-1044
- Saccharomyces cerevisiae, complementary DNA, genes, messenger RNA, molecular weight, phenol, ribosomal RNA, transfer 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 simpleness, the method is further advantagous in terms of safety (omitting hazardous phenol) and cost efficiency.