Isolation of plasmid DNA Sample Clauses

Isolation of plasmid DNA. Single E. coli colonies were used to inoculate 5 mL LB cultures, which were then grown shaking overnight at 250 rpm and 37°C. Plasmid DNA was isolated using the QIAprep Spin Miniprep Kit (QIAGEN) and DNA concentration was estimated using the Nanodrop (ND-1000 spectrophotometer, Thermo Scientific) as described in 2.8.2.
Sample 1
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Isolation of plasmid DNA. Miniprep Bacterial clones were selected for mini-culture and submerged in 4mL LB containing ampicillin. The bacterial culture was shaken at 37°C at 220rpm for 16 hours. After scaling up the bacterial culture the cells were pelleted. Preparation of DNA from minipreps was completed via the Miniprep kit (Qiagen). Analytic digestion with restriction enzymes and analysis of fragment size by agarose gel electrophoresis identified clones positive for the CAR construct. Sequencing (SourceBioscience) of positive clones was used to confirm successful cloning. A correct bacterial clone was then inoculated into 200mL LB with ampicillin and shaken at 37°C at 220rpm overnight. DNA was isolated from bacteria using the Maxiprep kit (Qiagen) following supplier’s instructions.
Sample 1

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