Qiagen
Plasmid Midi Kit
12143
Info: View Product View Product Specs
We used this kit to purify our plasmid of interest to transfect into mammalian cell lines. This kit provides a high quality plasmid with impressive concentration which can then be used to transfect into mammalian cell lines.
Gain of function assay after transfecting the gene of interest
Bacterial cell culture
25 ml of bacterial culture was harvested at 6000g and the pellet was resuspended in p1 buffer. We then added p2 and mixed vigorously. To the above reaction mixture, p3 buffer was added and incubated on ice and centrifuged at 15000 rpm and the supernatent is allowed to pass in through eqlibrated Qiagen tip. The DNA was eluted using QF buffer and the DNA is precipitated using isopropanol. The pellet is then washed with 70% ethanol and the pellet is re-suspended in water and quantified.
Use 100 ml baterial culture for a low copy number plasmid.
A very high quality plasmid is obtained.
None
High quality plasmid is obtained in very little time.
Cost is a bit high
I would recommend use of this kit to obtain a trasfection grade plasmid.
Product Name: QIAGEN Plasmid Midi KitSupplier Page