SAMN01977956
External Id
UCLA
INSDC center name
2013-03-14T00:00:00Z
INSDC first public
2021-02-28T02:04:57.515Z
INSDC last update
SRS418228
INSDC secondary accession
live
INSDC status
Generic
NCBI submission model
Generic.1.0
NCBI submission package
SRS418228
SRA accession
2011-05-31
collection date
Random mutagenesis was performed with N’-nitro-N-nitrosoguanidine (NTG). Briefly, cultures of ppc- were grown to exponential phase in LB medium, washed twice with 0.1M citrate buffer and then concentrated two-fold by centrifugation and suspension in 0.1 M citrate buffer (pH 5.5). Samples of 2mL were exposed to NTG at a final concentration of 50 mg/ml for 30 minutes at 37°C to reach a percentage kill of approximately 50%. The cells were washed twice with 0.1 M phosphate buffer (pH 7.0) and grown in LB for one hour. The cells were then challenged by plating on glucose minimal medium and grown at 37°C for 3 days. This procedure was performed on 24 separate samples of ppc-, each of which was plated separately to ensure genetically distinct populations of mutants. One colony from each separate NTG experiment was selected, restreaked on selective medium plates to verify the phenotype and then cultured in liquid medium to obtain genomic DNA.
description
USA: Los Angeles, CA
geo loc name
Escherichia coli JCL16
http://purl.obolibrary.org/obo/NCBITaxon_1286168
organism
JCL16
strain
24 NTG-derived and screened mutants of parent strain JCL16, sequenced in pooled libraries.
title