ERC000011
ENA-CHECKLIST
2023-12-01T00:42:22Z
ENA-FIRST-PUBLIC
2023-12-01T00:42:22Z
ENA-LAST-UPDATE
SAMEA112622028
External Id
School of Biosciences University of Birmingham
INSDC center name
2023-12-01T00:42:22Z
INSDC first public
2023-12-01T00:42:22Z
INSDC last update
public
INSDC status
ERS14614044
SRA accession
E-MTAB-12627:Rob N-terminal 3xFLAG tag sample 2
Submitter Id
ArrayExpress
broker name
Protocols: Salmonella enterica subsp. enterica serovar Typhimurium SL1344 is a histidine auxotroph of parental strain ST4/74. Hoiseth,S.K. and Stocker,B.A.D. (1981) Aromatic-dependent Salmonella typhimurium are non-virulent and effective as live vaccines. Nature, 291, 238-239. Cells were grown in liquid LB minimal medium at 37 degrees with shaking. Immunoprecipitations were done as described by Haycocks et al. (PLoS Pathog. 2015. 11:e1004605) using Vibrio cholerae strain E7946. After growing cells to midlog phase, crosslinking, and sonication, protein DNA complexes were immunoprecipitated using anti-FLAG or anti-Myc. 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.
description
pAMNFrob
genotype
mid exponential phase
growth condition
not applicable
isolate
Salmonella enterica subsp. enterica serovar Typhimurium str. SL1344
organism
Salmonella enterica subsp. enterica serovar Typhimurium str. SL1344
organism
nucleoid
organism part
Salmonella enterica subsp. enterica serovar Typhimurium str. SL1344
scientific_name
Rob N-terminal 3xFLAG tag sample 2
title