Maxi Fast Ion Endotoxin Free Plasmid Kit - 25 Preps
- Sample Size: 100ml high-copy plasmid/250ml low-copy plasmid
- Operation: Gravity Flow Column
- Expectant Yield: 500µg to 1mg of plasmid DNA
- Binding Capacity: 1.5mg
- Operation Time: 120 minutes or less
The Maxi Fast Ion Endotoxin Free Plasmid Kit uses pre-packed ion-exchange resin columns to purify plasmid or cosmid DNA from bacterial cultures. In this process, the modified alkaline lysis method and RNase treatment are used for creating cleared lysate with minimal genomic DNA and RNA contaminants. Using a gravity-flow procedure, the plasmid DNA in crude lysate has been bound to the column. The contaminants can be washed off with a wash buffer. Finally, the purified DNA is eluted by a high salt buffer and then precipitated with isopropanol for desalting. The entire procedure can be completed in less than two hours. The purified plasmid DNA is suitable for transfection, sequencing reactions, PCR, and in-vitro transcription.
The quality of the Maxi Fast Ion Endotoxin Free Kit is tested on a lot-to-lot basis by isolating plasmid DNA from a 100 ml overnight E. coli (DH5α) culture, containing pBluescript (A600>2U/ml). More than 400 µg of plasmid DNA is quantified with a spectrophotometer. The purified plasmid (1 µg) is used in EcoRI digestion, and checked by electrophoresis.
|Specifications||Maxi Fast-Ion Plasmid Kit (Endotoxin-Free)|
|Binding Capacity:||1.5 mg|
|Endotoxin Level:||<0.10 EU/µg|
|Sample Concentration:||200-1600 ml|
|Sample Type:||cultured bacterial cells (high-copy = 200-800 ml, low-copy = 500-1600 ml)|
|Fragment Size:||1-20 kb|
|Typical Yield:||1-1.5 mg|
|Elution Volume:||500 µl- 2 ml|
|Operation Time:||<180 minutes|
What competitor’s kit is comparable? Qiagen EndoFree Plasmid Maxi Kit
What are the new starting volumes and yields? The protocol has been revised as such: the maxi plasmid kits yield up to 1.5 mg of transfection grade plasmid DNA from 400 ml of cultured bacterial cells.
What is the endotoxin level of eluted DNA in the Maxi Fast-Ion Plasmid Kit? The endotoxin level is <0.1 EU/µg DNA. This level was determined by LAL testing.