title: Recovery of total microbial RNA from lactic acid fermented foods with a high starch content.
abstract: An optimized procedure for the recovery of RNA from micro-organisms involved in the fermentation of starchy foods (mainly hard-to-lyse lactic acid bacteria) is reported. Critical steps for the extraction were: cell recovery by differential centrifugation; cell wall digestion with both mutanolysin and lysozyme; and CTAB treatment for the elimination of starch. Digestion of starch with alpha-amylase did not improve extraction yields. The method yielded high amounts of RNA from pozol, a Mexican maize-based fermented food, and was found to extract total RNA efficiently from all the micro-organisms potentially present in these ecosystems. Both rRNA and mRNA recovered were of high quality and suitable for hybridization studies.
authors.affiliation: Institut Français de Recherche pour le Développement en Coopération (ORSTOM), Laboratoire de Biotechnologie Microbienne Tropicale, Montpellier, France. Frederic.Ampe@mpl.orstom.fr
authors.forename: F
authors.initials: F
authors.lastname: Ampe
authors.forename: N
authors.initials: N
authors.lastname: ben Omar
authors.forename: J P
authors.initials: JP
authors.lastname: Guyot
mesh_terms: D001419:Bacteria / Q000187:drug effects / Q000235:genetics; D015152:Blotting, Northern; D004587:Electrophoresis, Agar Gel; D005516:Food Microbiology; D005658:Fungi / Q000187:drug effects / Q000235:genetics; D007778:Lactobacillus / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification*; D009693:Nucleic Acid Hybridization; D012329:RNA, Bacterial / Q000302:isolation & purification*; D012331:RNA, Fungal / Q000302:isolation & purification; D013213:Starch
chemical_list: D012329:RNA, Bacterial; D012331:RNA, Fungal; D013213:Starch