Thermo Fisher Scientific
One Shot ccdB Survival 2 T1R Competent Cells
A10460
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We transformed the one shot ccdB competent cells with PLEX305 lentiviral plasmid (50ng) obtained from addgene into ccdB competent cells. Incubated on ice for 30 mintues. Heat shock at 42 C for 30 sec. Kept on ice for 5 minutes. Added 25ouL of SOC medium provided with kit and incubated at 30C for one hour. Plated 20uL of cells on LB+Ampicillin plates overnight at 30C. Observed colonies were inoculated in to 5mL LB medium with ampicillin. Minipreps were done with Qiagen kit and isolated plasmid was ran on agarose gel for size confirmation and digested with restriction enzymes for conformation of plasmid.
Amplification and isolation of lentiviral plasmid
Lentiviral plasmid and ccdB competent cells
For cloning unstable inserts, incubate cells at 30 C
We observed colonies after 24 h of incubation at 30c. Plasmids prepared from these colonies were confirmed by sequencing and digestion
N/A
Efficient transformation
Excellent competent cells for transformation of lentiviral ccbB gene expressing plasmids