Cell lines were transfected with pEZ-LV206-PANDAR, pEZ-LV206-control and extracted with Plasmid Midiprep System, Promega. PC9 and A549 cells were cultured for 24 h and transfected into six-well plate using Lipofectamine3000. The transfected cell lines were harvested after 48 h or 72 h for follow-up series of cell assays. Besides, the experimental methods of two plasmids co-transfection were same as above.
Plasmid midiprep system
Manufactured by Promega
Sourced in Germany
The Plasmid Midiprep System is a laboratory equipment product designed for the purification of plasmid DNA from bacterial cultures. It is a automated system that efficiently extracts and purifies plasmid DNA for various applications in molecular biology and genetic research.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using plasmid midiprep system
1
Plasmid Transfection and Cell Assays
2
TcC Variant Mutagenesis and Expression
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All DNA manipulations described in this work were performed by standard methods.41 Megaprimers for the TcC variants, possessing the specific mutation (Table S1† ) were purchased from Microsynth AG (Switzerland) and used in two-stage PCR reactions to introduce mutations using Pfu DNA polymerase (Promega GmbH, Germany) and pET26b(+)_TcC as template. Restriction enzymes were purchased from New England Biolabs and used following the manufacturer's protocol. Plasmid DNA was prepared using PureYield™ Plasmid Miniprep and Plasmid Midiprep system (Promega GmbH, Germany). The amplified products were chemically transformed in Escherichia coli XL-10.
For HiC the codon-optimized genes (i.e. E. coli codon usage) of wild-type, I167Q and L64H/I167Q variants were purchased from GeneArt® (Life Technologies, USA). The genes were cloned into the vector pET26b(+) and chemically transformed by heat shock into E. coli XL-10.
For HiC the codon-optimized genes (i.e. E. coli codon usage) of wild-type, I167Q and L64H/I167Q variants were purchased from GeneArt® (Life Technologies, USA). The genes were cloned into the vector pET26b(+) and chemically transformed by heat shock into E. coli XL-10.
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