title: Extraction of RNA from Ca-alginate-encapsulated yeast for transcriptional profiling.
abstract: We have developed a method for preparing high-quality total RNA from Ca-alginate-encapsulated Saccharomyces cerevisiae that is suitable for microarray analysis. Encapsulated cells were harvested from immobilized cell reactors and flash-frozen in liquid nitrogen. Following low-temperature mechanical disruption, cells were freed from Ca-alginate by reverse ionotropic gelation and purified by centrifugation, and then total RNA was extracted using hot acid phenol. The yield and quality of the RNA were consistently high; the RNA was free of contaminating alginate, and in microarray analysis it performed as well as RNA isolated from planktonic cells.
authors.affiliation: Division of Biological Sciences, University of Montana, Missoula, MT 59812, USA.
authors.forename: Arthur L
authors.initials: AL
authors.lastname: Kruckeberg
authors.forename: Saisubramanian
authors.initials: S
authors.lastname: Nagarajan
authors.forename: Kate
authors.initials: K
authors.lastname: McInnerney
authors.forename: Frank
authors.initials: F
authors.lastname: Rosenzweig
mesh_terms: D000464:Alginates / Q000737:chemistry*; D002118:Calcium / Q000737:chemistry*; D020869:Gene Expression Profiling; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D012331:RNA, Fungal / Q000302:isolation & purification*; D012441:Saccharomyces cerevisiae / Q000235:genetics*
chemical_list: D000464:Alginates; D006603:Hexuronic Acids; D012331:RNA, Fungal; D020723:Glucuronic Acid; D002118:Calcium