Modern plasmid DNA separation

Modern plasmid DNA separation and purification of separation from escherichia coli, since big represented in e. (E.c oli) and molecular biology, from the important status in the E.c oli separation and purification qc Piasmid < > become DNA DNA in recent years over an important topic of centrifugal technology. But the plasmid DNA separation and purification of super fast and centrifugal equipment (speeding centrifuge, turn and accessory) put forward higher request.
E.c oli is typical of prokaryote cells, due to lack of its nuclear prokaryote cells that have cell membranes of the unit can be separated with multiple functions for the components of the regional and local independent system, thus not endometrial cells contain nuclei (nuclear, organelles endoplasmic reticulum, golgi apparatus, body, the lysosome, etc.). Electron micrograph E.c oli two can show the difference between internal areas -- the cytoplasm and nuclear, in their around a layer of thinner outside the plasma membrane and very thick in the cell walls, and some end external attached of free flagellum. Plasmid DNA located in nuclear zone, filaments exist, such filaments in many cases is material very long annular DNA fragments of the secret that folded.
On the microstructure E.c oli to point, the super centrifugal separation and purification plasmid DNA sequence is before the pretreatment of:
E.c oli to use lysozyme, surfactants used to cell Trit, such as SDS X - 100, acetic acid, etc EE membrane pot that DNA, RNA and protein most sedimentation (90%).
Sediment can add TE buffer 10m (MTris - lmMEDTA pH8.0), HCL in - after the protein, and advances to go RNA, Can also be used to protein, speed centrifugation (level, to go to the RNA DNA or DNA fragments.
The plasmid DNA separation method of speeding.
The traditional separation method: a few years ago by equipment, plasmid DNA restriction of general use CsCl balance, separated from forming gradient centrifugation, density. With 10 ~ 12ml single capacity, for example, in turn, sling flat 36.000 RPM separated by 60 hours with Angle type head, 45, separation, the former inter OOOrpm 36 hours, including deceleration share to 1.3 billion of life, the latter to drive to use to 1 million to drive, the ministry of life at speed centrifuge total lifespan of 100 to 200 million each turn, undoubtedly, plus high cost experiment, the price of your CsCl dosage, make this type of factors such as the separation and purification of experimental work become very expensive.