Affymetrix GeneChip Yeast Genome S98 [YG_S98]

Phenomic and transcriptomic analyses reveal that autophagy plays a major role in desiccation tolerance in Saccharomyces cerevisiae

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Response of Saccharomyces cerevisiae strain BY4741 to desiccation

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Transcription profiling by array of yeast desiccation stress response in a time series

Transcription profiling by array of yeast desiccation stress response to dessication in a time series

Response of Saccharomyces cerevisiae strain BY4741 to desiccation


1. Resuspend filtered cells in 400 μL of TES buffer, and  add 400μL acid phenol. Vortex for 10 sec, then incubate for ~60 min at 65oC in a shaking heating block. 2. Incubate on ice for 5min. Microfuge for 5 min at 4oC. 3. Transfer the upper aqueous layer to a new 1.5 ml tube, and add 400μL fresh acid phenol. Repeat step 2. 4. Add 200microL TE to the recovered aqueous layer sample, and extract once with 400 μL chloroform; vortex vigorously, spin 5 min , 4oC, and take the upper aqueous layer for further use. 5. Precipitate the RNA from the sample as follows. Add 10% volume of 3M NaOAc, pH5.2 and 2-2.5 volume of 100% EtOH. Incubate on ice for at least 1 hour, then pellet RNA by microfuging at full speed for 15 min, 4oC. 6. Remove the supernatant, and wash the pellet with 500μL 80%EtOH. Allow the pellet to air dry. 7. Resuspend the pellet in 50μL of RNase-free water. Store at -70oC.(Parameters: Extracted product = total_RNA, Amplification = none).

A 2 ml volume of culture was filtered through 0.45μm cellulose nitrate membrane filters (Whatman) and dried in an incubator at 30oC for 0, 30, 60 and 120 min.

CEL files were normalised using RMA, as implemented in the R/Bioconductor suite of tools

Strain BY4741 was grown to late post-diauxic phase (A600 ~ 3.0) in YEPD at 30oC with shaking at 160 rpm, taking care not to allow cells to reach true stationary phase before being used.(Parameters: start time = 0, stop time = 48, time unit = hours, min temperature = 30, temperature unit = C, media = YEPD)

Transcription profiling by array of yeast desiccation stress response in a time series

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