Illumina HiSeq 2500 sequencing; Genome-wide analysis reveals that budding yeast Rif1 binds to replication origins and at blocked replication forks to protect nascent DNA
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
No description
Antigen
NA
Cell type
Cell type Class
No description
Cell type
NA
NA
NA
Attributes by original data submitter
Sample
ENA first public
2018-08-16
ENA last update
2018-04-30
ENA-CHECKLIST
ERC000011
External Id
SAMEA4623656
INSDC center alias
Centre for Genome Enabled Biology and Medicine University of Aberdeen
INSDC center name
Centre for Genome Enabled Biology and Medicine University of Aberdeen
INSDC first public
2018-08-16T17:02:56Z
INSDC last update
2018-04-30T13:36:02Z
INSDC status
public
Submitter Id
E-MTAB-6736:2017_006B_4
broker name
ArrayExpress
common name
baker's yeast
genotype
Rif1-13myc
growth condition
normal S phase, 60 minutes
sample name
E-MTAB-6736:2017_006B_4
Sequenced DNA Library
library_name
2017_006B_4_s
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Collected cells were fixed with formaldehyde at room temperature, washed with TBS buffer and resuspended in lysis buffer. A multibead shocker was used to disrupt the cells and chromatin was sheared by sonication. Finally antibodies bound to magnetic Dynabeads were added and incubated. All cells were cultivated in YPD. Alpha-factor arrested cells were treated for two hours with alpha factor at 30 degrees celsius. HU-arrested cells were first synchronised in alpha factor and then released in to YPD containing 0.2 M hydroxyurea and cultivated for 60 minutes. Normal S Phase cells were arrested by alpha factor at 16 degrees celsius and released into fresh YPD medium at 16 degrees celsius, and then collected at 60 and 90 minutes. Chromatin immunoprecipitation was performed using monoclonal anti-Myc antibody and Dynabeads Protein G. Precipitated DNA was cleaned used TE buffer, glycogen and Proteinase K before extraction with sodium chloride and phenol/chloroform/iso-amylalcohol. DNA was washed using ethanol and purified with a Qiagen PCR purification kit. Library construction using NEBNext Ultra II DNA Library Preparation Kit.