title: Fkh1-myc log replicate 2
organism: Saccharomyces cerevisiae
description: Protocols: 1. Grow on YPD+G418 plates 2. Pick colony mid-day into 75ml 1/2 diluted delft+CSM+2%glu 3. Next morning, start mid-log phase culture, 4x 250ml shakeflasks with 150ml 1/2 diluted delft+CSM+2%glu. Add to make starting OD 0.2. Leave ~5h before checking OD. Target 0.8. ON cultures are left shaking until afternoon. ChIP-seq on beads: Pierce Anti-c-Myc Magnetic Beads (88843). 1. From OD, calculate how to get 3x33 OD into 3x50ml tubes. Prepare water with 37% formaldehyde 1:36, 1.4ml to each 50ml tube then add appropriate amount of cultures to get 33OD. Shake on tipping board RT 15min 2. Add 2.5M glycine to a final concentration of 125mM (1:19), 2.6ml to 50ml tubes, leave on shaker 5min 3. Pellet cells 1500g 4c 3min. Wash once in 20ml then once in 10ml ice-cold TBS 4. After final wash, move to eppi with small remaining amount of liquid. Get box with liquid N2 and drop tubes in. Move to -80c for storage ChIP-exo library preparation as described in manuscript
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2019-09-30T04:05:11Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-10-22T14:44:16Z
attributes.tag: External Id
attributes.value: SAMEA5040253
attributes.tag: INSDC center name
attributes.value: Department of Biology and Biological Engineering, Chalmers University of Technology
attributes.tag: INSDC first public
attributes.value: 2019-09-30T04:05:11Z
attributes.tag: INSDC last update
attributes.value: 2018-10-22T14:44:16Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-7323:Fkh1-myc log replicate 2
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: baker's yeast
attributes.tag: genotype
attributes.value: MATa his31 leu20 met150 ura30 FKH1-MYC9::kanMX6
attributes.tag: growth condition
attributes.value: logarithmic growth phase
attributes.tag: sample name
attributes.value: E-MTAB-7323:Fkh1-myc log replicate 2
attributes.tag: scientific_name
attributes.value: Saccharomyces cerevisiae
attributes.tag: strain
attributes.value: BY4741