Illumina MiSeq sequencing; ChIP-exo of S.cerevisiae transcription factors Fkh1 and Fkh2 under logarithmic growth and stationary
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
No description
Antigen
NA
Cell type
Cell type Class
Yeast strain
Cell type
BY4741
NA
NA
Attributes by original data submitter
Sample
ENA first public
2019-09-30
ENA last update
2018-10-22
ENA-CHECKLIST
ERC000011
External Id
SAMEA5040253
INSDC center alias
Department of Biology and Biological Engineering, Chalmers University of Technology
INSDC center name
Department of Biology and Biological Engineering, Chalmers University of Technology
INSDC first public
2019-09-30T04:05:11Z
INSDC last update
2018-10-22T14:44:16Z
INSDC status
public
Submitter Id
E-MTAB-7323:Fkh1-myc log replicate 2
broker name
ArrayExpress
common name
baker's yeast
genotype
MATa his31 leu20 met150 ura30 FKH1-MYC9::kanMX6
growth condition
logarithmic growth phase
sample name
E-MTAB-7323:Fkh1-myc log replicate 2
strain
BY4741
Sequenced DNA Library
library_name
Fkh1-myc log replicate 2_s
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
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