characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2017-08-24T17:01:09Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2017-08-14T10:49:25Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA104212279
characteristics.characteristic: External Id
characteristics.text: The Wadsworth Center, NYSDOH
characteristics.characteristic: INSDC center name
characteristics.text: 2017-08-24T17:01:09Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-08-14T10:49:25Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS1871297
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-5969:Plasmid loss cas3-1
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: Protocols: • Start a 2 mL overnight for each Plasmid loss strain ( 13 different protospacer plasmids) in LB + Kan and amp at 37C, 225rpm. • Subculture all strains together into 10 mL of LB (12.5 L for 8 protospacers and 7.7 L for 13 protospacers) • Make electrocomp cells • Electroporate in pBAD33 and pBAD33-Cas3 WT into strains • Plate onto M9 + 0.2% glycerol + 0.2% arabinose + chloramphenicol carbenicillin • Next day: Scrape plates into 1.5 mL of LB, spin down, dump supernatant, and can freeze at -20 C here. pelleted cells were used as the source of DNA for PCR amplification o PCR protospacer with ultramers using a tiny bit of cells 40 L Taq Rxn, 68 anneal, 30 sec extension• Fwd: 1:10 dilution of 8567• Rev: 1:100 dilution of 8556-8566 and 8537
characteristics.characteristic: description
characteristics.text: cas3::PcseI cas1 yggX::kan + pBAD33-wt cas3
characteristics.characteristic: genotype
characteristics.text: Escherichia coli str. K-12 substr. MG1655
characteristics.characteristic: organism
characteristics.text: Escherichia coli str. K-12 substr. MG1655
characteristics.characteristic: organism
characteristics.text: 1
characteristics.characteristic: replicate
characteristics.text: Escherichia coli str. K-12 substr. MG1655
characteristics.characteristic: scientific_name
characteristics.text: Plasmid loss cas3-1
characteristics.characteristic: title