Sigma-Aldrich
GenElute™ HP Plasmid Midiprep Kit
NA0200-1KT
I do work with liver pathology and have a strong background and experience in innovative liver tumor models, molecular biology techniques such as cloning, mutagenesis and gene expression along with cell and tissue culture. I also have extensive experience in working with transgenic and knockout mouse models and have used sleeping beauty transposon technology along with hydrodynamic delivery method to generate tumors in mouse livers that are composed of human cancer causing genes (oncogenes). We use this kit as it is very convenient for doing midi-preparation of plasmid from bacterial culture, the whole process takes about 30 minutes, the protocol should be: 50 ml culture media for 1 column, but it could be used for 100-150ml media culture. Additionally, the preparation doesn’t need phenol/chloroform extraction or alcohol precipitation operation. In general, we save much more time with the kit, and people can also prepare vacuum instrument if they don’t want to buy special vacuum instrument.
Plasmid midi-preparation
Followed protocol
We just followed the protocol book, 4ml re-suspend solution, 4 ml lysis buffer, then mix and lyse for 5 minutes, add with 4 ml neutral buffer and 3 ml binding buffer, mix well and let stand for 5 minutes, then filter by syringe and transfer the column preparation buffer treated column, after going through, then wash with washing buffer 1 and buffer 2.
Don't shake vigorously
VERY nice kit, and it can save many time compared to other kits
None
Works well
One of the instruments was constructed by me, since we didn't have the vacuum machine