Sigma 38 ChIP-seq replicate 1
Escherichia coli K-12
Protocols: Bacterial strain RPB104 is a laboratory strain of Escherichia coli Cells were grown in liquid LB medium at 37 degrees with shaking Immunoprecipitations were done as described by Haycocks et al. (PLoS Pathog. 2015. 11:e1004605) using bacterial strain RPB104 delta hns. After first growing cells to stationary phase, crosslinking, and sonication, protein DNA complexes were immunoprecipitated using anti-FLAG (Sigma). Libraries were prepared using immunoprecipited protein-DNA complexes immobilised with Protein A-sepharose. DNA fragments were then blunt ended, A-tailed and barcoded. This was done using an NEB quick blunting and ligation kit, the Klenow fragment (5'-3' exo-, NEB) and NEXTflex ChIP-seq barcodes (Bioo Scientific). Following elution of complexes from the Protein-A sepharose, crosslinks were reversed and barcoded libraries were amplified by PCR. The number of PCR cycles was determined empirically for each library. After amplification, library concentration was quantified using Qubit analysis and real time PCR.
ENA-FIRST-PUBLIC
2021-03-01T12:15:51Z
ENA-LAST-UPDATE
2020-02-11T10:31:46Z
External Id
SAMEA6534363
INSDC center name
University of Birmingham
INSDC first public
2021-03-01T12:15:51Z
INSDC last update
2020-02-11T10:31:46Z
INSDC status
public
Submitter Id
E-MTAB-8778:Sigma 38 ChIP-seq replicate 1
broker name
ArrayExpress
developmental stage
stationary phase
genotype
hns::kan rpoS-SPA
organism part
whole organism
sample name
E-MTAB-8778:Sigma 38 ChIP-seq replicate 1
sampling site
bacterial nucleoid
scientific_name
Escherichia coli K-12
strain
RPB104