FavorPrep™ Plasmid DNA Maxi Extraction Kit (Endotoxin free)

FavorPrep™ Plasmid DNA Maxi Extraction Kit (Endotoxin free)詳細資料

Description

The FavorPrep&amptrade Plasmid Extraction Midi and Maxi Kits provide a pre-packed ion-exchange resin column to purify plasmid or cosmid from 50-250ml bacterial cultures. When using the standard protocol, the entire process can be completed in 120 minutes or less and up to 500&ampmug of plasmid can be expected. The purity of purified plasmid DNA is equal to that obtained by 2x CsCl-gradient centrifugation and suitable for all molecular biological applications.

 

Features

★            Purity: equal to that obtained by 2x CsCl-gradient centrifugation.

★            Safe: Eliminates the use of phenol, chloroform, ethidium bromide, and cesium chloride, minimizing exposure to, and disposal of hazardous materials.

★            Time saving: Complete the process in less than 120 minutes.

 

Applications

The purified plasmid DNA can be immediately used in any downstream molecular biology application.

★          Transfection

★          Microinjection

★          Sequencing

★          PCR

★          Restriction enzyme digestion

 

Operation Time

Less than 120 minutes

 

Operation Format

Gravity-Flow

 

Agarose Gel Analysis of Plasmids Purified

 

 

 

 

 

 

 

 

 

 

Agarose gel analysis of various plasmid DNA purified with Plasmid DNA Midi/Maxi Kit

C: crude lysate

F: flow-through

W: wash

E: elute

M: 1Kb DNA Ladder

Plasmid pBluescript (high-copy) and pBR322 (low-copy) are purified with the Plasmid DNA Midi/Maxi Kit.  DNA in crude lysate from alkaline lysis and fractions from each gravity-flow step was collected by isopropanol precipitation and load to each lane.

 

 

Recommended  Culture Volume      Expected Yield

High-copy plasmid     Low-copy plasmid         

 

       Midi    50 ml            100ml              up to 200&ampmug

 

       Maxi   100ml            250ml              up to 500&ampmug

 

 

Procedure

 

In the process, the modified alkaline lysis method and RNase treatment are used to get cleared cell lysate plasmid DNA.  The plasmid DNA has been bound to the column, the contaminants can be washed off with wash buffer.  Finally, the purified plasmid DNA is eluted by a high salt buffer and then precipitated with isopropanol for desalting.  The entire procedure can be completed in 120minutes without ultracentrifuges and HPLC or other toxic reagents. 

 

 

  

 

 

 

 

 

 

 

 

Storage Conditions

Plasmid DNA Extraction Midi/Maxi Kit can be stored at room temperature (15-25℃).  After adding RNase A, PM1 Buffer, should be stored at 4℃ and is stable for six months.  Other buffers and columns can be stored dry for up to 1 year at room temperature (15-25℃).