When used for the first time, add all RNase A carried by the kit to Buffer P1 (final concentration 100ug/ml) and store it at 4℃. If RNase A is inactivated in Buffer P1, the extracted plasmid may be mixed with trace RNA residues. Add RNase A to Buffer P1.
When the ambient temperature is low, SDS in Buffer P2 may precipitate and appear turbid or precipitated. It can be heated in a 37℃ water bath for a few minutes to restore clarity. Do not shake violently to avoid excessive foam formation.
Avoid volatilization, oxidation and pH change of reagents exposed to air for a long time, and close the cover of each solution in time after use.
Buffer ER is transported at room temperature. It can be stored at 4℃ for one month, and stored at -20℃ for a long time.
Components
Component
Storage
PLK1301 10 Preps
RNaseA(10mg/ml)
-20℃
750µl
Buffer P1
4℃
77 ml
Buffer P2
RT
77 ml
Buffer N3
RT
77 ml
Buffer ER
-20℃
25 ml
Buffer PE
RT
63 ml
Buffer WB
RT
25 ml×2
Buffer EB
RT
20 ml
Adsorption column DC
RT
10
Collection pipe (50ml)
RT
10
Description
This kit uses an improved SDS alkaline lysis method to lyse cells. The crude extract is selectively combined with centrifugation to remove endotoxin through a unique buffer Er, and then the silicon matrix membrane in the centrifugal adsorption column selectively binds plasmid DNA in the solution under high salt and low pH, and then removes impurities and other bacterial components through buffer WB. Finally, the pure plasmid DNA is eluted from the silicon matrix membrane by buffer EB with low salt and high pH.
Features
The silicon matrix membranes in the centrifugal adsorption column are all specially made adsorption membranes imported from world-famous companies, with minimal difference in adsorption capacity between columns and good repeatability. It overcomes the disadvantage of unstable quality of domestic kit membrane.
There is no need to use toxic phenol, chloroform and other reagents, nor ethanol precipitation. It is fast and convenient. 0.5-2mg of pure high copy plasmid DNA can be rapidly extracted from 150-300 ml of Escherichia coli lb (Luria BERTANI) culture solution, and the extraction rate is 80-90%.
The unique process formula can remove endotoxin, and the endotoxin content is very low (<0.1eu/μg DNA), and the cell transfection effect was excellent. It can also be directly used in enzyme digestion, transformation, PCR, in vitro transcription, sequencing, and other molecular biology experiments.