2024-05-06
ENA first public
ERC000011
ENA-CHECKLIST
The Wadsworth Center, NYSDOH
INSDC center name
public
INSDC status
ERS19224922
SRA accession
E-MTAB-14044:∆cas1-4-rep2
Submitter Id
ArrayExpress
broker name
not collected
collection date
Protocols: 1.0 µL from each pellet was resuspended in 20µL dH2O and incubated at 95˚C for 10 minutes.    PAM libraries were transformed into E. coli S17.  E. coli S17 + PAM libraries were grown in LB with 30 µg/mL Chloramphenicol to OD600 1.0, and then pooled equally.  The E. coli  S17 +PAM libraries were introduced into V. cholerae strains by conjugation, as previously described (Box et al., 2015).  100µL of cells from each conjugation were plated on LB plates containing 75 µg/mL kanamycin and 2.5 µg/mL chloramphenicol.  The plates were scraped the next day, and cells were resuspended in LB and pelleted.  From each cell resuspension, 1.0 µL was used as a template in a PCR reaction with universal forward oligonucleotide and reverse index oligonucleotides.  From each cell resuspension, 1.0 µL was used as a template in a PCR reaction with universal forward oligonucleotide and reverse index oligonucleotides.
description
KS808 CRISPR/Cas::KanR Δcas1
genetic modification
not collected
geographic location (country and/or sea)
not applicable
isolate
Vibrio cholerae
organism
Vibrio cholerae
organism
whole organism
organism part
protospacer4
pam library plasmid
Vibrio cholerae
scientific_name
∆cas1-4-rep2
title