HiYield Plasmid Mini Kit 3.0 is specially designed for rapid isolation of plasmid or cosmid DNA from 1-4 ml of bacterial cultured cells. As high as 50 µg of high-quality plasmid DNA can be purified in less than 30 minutes and is ready for restriction digestion, ligation, PCR, and sequencing reaction. No phenol extraction or alcohol precipitation is required in this protocol. In the process, clear and extra pure cell lysate with minimal genomic DNA and RNA contaminants can be obtained through the modified alkaline lysis method and RNase treatment. In the presence of a chaotropic salt, the plasmid DNA within the lysate will then bind to the glass fiber matrix equipped in the spin column. The contaminants are washed away with an ethanol-containing wash buffer. Finally, the purified plasmid DNA is eluted by a low salt elution buffer or distilled water. Typical yields of high-purity are 20~50 µg for high-copy number plasmids or 3~10 µg for low-copy-number plasmids.
HiYield Plasmid Mini Kit 3.0 improves the structure of the membrane and the composed of glass fiber makes the purified plasmids purer and the lower concentration of endotoxin. 

• Sample :
  1 – 4 mL of bacterial cells

• Format :
  spin column (centrifuge)

• Yield :
  up to 50 μg of plasmid/cosmid DNA

• Operation time :
  up to 30 minutes

• Elution volume :
  50 – 100 μL

Restriction enzyme digestion, library screening, ligation, PCR, transformation/sequencing reactions

The quality of HiYield Plasmid Mini Kit 3.0 is tested on a lot-to-lot basis. The kit is tested by isolation of plasmid DNA from 5 ml culture of E.coli DH5α transformed with the plasmid pGAD424 (A₆₀₀> 2 units/ml). More than 40 µg of plasmid DNA should be obtained. One µg of the purified product is also tested for restriction enzyme digested with EcoR I followed by agarose gel analysis.