title: Treated with 50 uM NaF
organism: Saccharomyces cerevisiae
description: Protocols: After growth, cells were spun down and washed twice with water. Cell count was determined by measuring O.D., and 1 mL of cells at O.D. 4 were transferred to fresh tubes. Four hours growth at 30C with shaking in 25 mL yeast extract peptone dextrose with adenine +/- 50 uM NaF The cells were resuspended in 200 μL AE buffer (50 mM NaOAc and 10 mM EDTA in water), 40 μL 10% SDS, and 200 μL 25:24:1 phenol:choloform:isoamyl alcohol (Sigma). Cell lysates were mixed and placed in a 65°C water bath for 10 minutes, vortexing cells at each minute point. The mixture was then placed on ice for 5 minutes, before being transferred to heavy phase lock gel tubes (5 Prime). The phase lock tubes were spun for 5 minutes at 14k rpm, after which 600 μL chloroform was added. The tubes were again spun, and the top fraction was placed into fresh 1.5 mL tubes. This fraction was resuspended in 1 mL pure ethanol and placed in a -20°C freezer overnight. In the morning, the mixture was spun down and the ethanol removed via evaporation. RNA was resuspended in sterile water, and the concentration established using a Nanodrop. 10 ug of RNA was treated for 20 minutes at 37°C with 1x DNase buffer and 1 uL Turbo DNAse (Invitrogen). Afterwards, RNA was again extracted using phenol:chloroform and ethanol. RNA was resuspended in sterile water and the concentration measured using a nanodrop. The S. cerevisiae library is publicly available at Ensemble, and was used as a reference.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2019-10-01T04:02:53Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2019-06-14T14:17:31Z
attributes.tag: External Id
attributes.value: SAMEA5718246
attributes.tag: INSDC center name
attributes.value: YALE UNIVERSITY
attributes.tag: INSDC first public
attributes.value: 2019-10-01T04:02:53Z
attributes.tag: INSDC last update
attributes.value: 2019-06-14T14:17:31Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-8061:Treated with 50 uM NaF
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: baker's yeast
attributes.tag: genotype
attributes.value: FEX1, FEX2 double knockout
attributes.tag: growth condition
attributes.value: cell suspension culture
attributes.tag: organism part
attributes.value: whole organism
attributes.tag: sample name
attributes.value: E-MTAB-8061:Treated with 50 uM NaF
attributes.tag: scientific_name
attributes.value: Saccharomyces cerevisiae
attributes.tag: strain
attributes.value: S288c