Extraction of RNA from Ca-alginate-encapsulated yeast for transcriptional profiling.
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.
Kruckeberg
Arthur L
AL
Division of Biological Sciences, University of Montana, Missoula, MT 59812, USA.
Nagarajan
Saisubramanian
S
McInnerney
Kate
K
Rosenzweig
Frank
F
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*
D000464:Alginates; D006603:Hexuronic Acids; D012331:RNA, Fungal; D020723:Glucuronic Acid; D002118:Calcium