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Yeast genomic expression patterns in response to zymolyase treatment

ABSTRACT: The evaluation of zymolyase treatment using yeast gene expression comparison analysis. The yeast BY4743 strain was used for this study. Yeast cells were treated with Zymolyase (final conc. 300 U/ml), and incubated at 37 degree for ten minutes. Digestion process was not provided for control sample. Total RNA was isolated with commercial kit (FastRNA Pro Red kit, Q-Biogene), and amplified RNA (aRNA) was synthesized with 3'IVT Express kit (Affymetrix). All samples were hybridized with Gene Chip (Yeast Genome 2.0 Array, Affymetrix), then each array chip was scanned by GeneChip Sanner 3000 (Affymetrix). Yeast were treated with zymolyase or left untreated. The only difference in conditions between the treated and untreated samples was the enzymatic digestion process. The conditioned samples were normalized with three replicates.

ORGANISM(S): Saccharomyces cerevisiae

SUBMITTER: Tadahiro Suzuki

PROVIDER: E-GEOD-36955 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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RNA preparation of Saccharomyces cerevisiae using the digestion method may give misleading results.

Suzuki Tadahiro T Iwahashi Yumiko Y

Applied biochemistry and biotechnology 20130117 5

Zymolyase (lyticase) is used for cell wall digestion in yeast experiments and is needed for incubation processes under moderate experimental conditions. This has been thought to cause unfavorable effects, and many researchers are aware that the enzyme method is unsuitable for RNA preparation following several reports of stress responses to the enzyme process. However, RNA preparation with enzyme digestion continues to be used. This may be because there have been insufficient data directly compar ...[more]

PMID: 23325148

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