Recovery of total microbial RNA from lactic acid fermented foods with a high starch content.
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.
Ampe
F
F
Institut Français de Recherche pour le Développement en Coopération (ORSTOM), Laboratoire de Biotechnologie Microbienne Tropicale, Montpellier, France. Frederic.Ampe@mpl.orstom.fr
ben Omar
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N
Guyot
J P
JP
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
D012329:RNA, Bacterial; D012331:RNA, Fungal; D013213:Starch