pCP20 vector map and sequence - novoprolabs.com
WEBpCP20 is an ampicillin and CmR plasmid that shows temperature-sensitive replication and thermal induction of FLP synthesis. CmR and KmR mutants were transformed with pCP20, and ampicillin-resistant transformants were selected at 30°C, after which a few were colony-purified once nonselectively at 43°C and then tested for loss of all antibiotic ...
One-step inactivation of chromosomal genes in Escherichia coli …
WEBJun 6, 2000 · pCP20 is an ampicillin and Cm R plasmid that shows temperature-sensitive replication and thermal induction of FLP synthesis . Cm R and Km R mutants were transformed with pCP20, and ampicillin-resistant transformants were selected at 30°C, after which a few were colony-purified once nonselectively at 43°C and then tested for …
ProcedureFLPFRTRecombination - Barrick Lab
WEBTransform with pCP20. This plasmid has a temperature-sensitive origin of replication, confers ampicillin resistance, and encodes the FLP recombinase. Add 500 µl SOC and incubate in a 1.7 ml tube shaking on its side at ≥120 rpm for 1 hr at 30°C to allow antibiotic resistance induction.
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Producing Gene Deletions in Escherichia coli by P1 Transduction …
WEBSep 1, 2018 · Propagate bacteria in a lysogeny broth (LB) medium 25 with vigorous shaking (180–200 rpm) at 37 °C or 30 °C in the case of BL21ΔABCF and strains containing pCP20. Perform plasmid curing at 43 °C. For a solid medium, supplement LB with 1% agar (w/v).
Site - pCP20 - Yale University
WEBE. coli Genetic Resources at Yale CGSC, The Coli Genetic Stock Center
SlyD-deficient Escherichia coli strains: A highway to …
WEBMay 23, 2018 · pCP20 is an Ap R and Cm R plasmid that exhibits temperature-sensitive replication and thermal induction of FLP synthesis [14]. Km R mutants were transformed using pCP20, and ampicillin-resistant transformants were selected at 30 °C.
CRISPR-FRT targets shared sites in a knock-out collection for off …
WEBJun 8, 2018 · A new pTarget plasmid encoding an sgRNA targeting a unique sequence on the recA plasmid could be transformed into the chromosomal recA library pool and plated on chloramphenicol + spectinomycin...
Genome modifications and cloning using a conjugally transferable ...
WEBAug 28, 2015 · We were able to use the Flp recombinase encoded on the temperature-sensitive plasmid pCP20 to successfully remove a FRT-flanked antibiotic resistance cassette used for genome modification in E. ictaluri.
pCP20 - СAGER
WEBpCP20. A plasmid for introducing deletions in E. coli genome. The plasmid carries gene coding for temperature-sensitive version of Rep101 protein needed for replication with the pSC101 origin.
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