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ISOLATE II Plasmid Mini Kit is designed for the rapid and efficient isolation of highly pure plasmid DNA from bacterial cultures using silica-membrane spin column technology. The isolation process combines alkaline lysis of the harvested bacteria, followed by clarification and subsequent specific binding of plasmid DNA directly to the filter of a spin column. Contaminants are efficiently removed during the washing steps. In the final step, highly-pure plasmid DNA is eluted. The kit shows excellent recovery of plasmid DNA and reproducible results even from low culture volumes . Separate protocols are provided for the isolation of high copy and low copy plasmids. In each case, intact, high purity plasmid DNA is isolated within 25 minutes. Typically, up to 25µg and 40µg plasmid DNA is obtained from 5ml and 10ml cultures, respectively. The isolated DNA shows excellent performance in downstream molecular biology applications such as PCR, transformation, cloning, sequencing , restriction analysis, etc. The online product manual has protocols for isolation of low-copy plasmids, P1 constructs and cosmid DNA from E. coli, isolation of plasmid DNA from Gram-positive bacteria as well as plasmid DNA clean-up. Frequent use of protocols for isolation of low-copy plasmids, P1 constructs and cosmids will require increased buffer volumes.

Features

• Rapid protocol: 25 minutes/18 preps
• High-capacity plasmid preparation kit
• High-purity plasmid DNA: typical A260/A280 ratio >1.8
• High-yields: up to 40µg from 10ml E. coli culture

Applications

• Isolation of high-copy plasmid DNA
• Isolation of low-copy plasmid DNA, P1 constructs and cosmids
• Isolation of plasmid DNA from Gram-positive bacteria
• Plasmid DNA clean-up
• Isolated DNA is ready for downstream applications such as PCR, real-time PCR, cloning and sequencing

Components: ISOLATE II Plasmid Mini Spin Columns Collection Tubes Resuspension Buffer P1 Lysis Buffer P2 Neutralization Buffer P3 Wash Buffer PW1 Wash Buffer PW2 Elution Buffer P RNase A (lyophilized)

Conditions: ISOLATE II Plasmid Mini Kit contents should be stored dry at room temperature. Under these conditions, the kit is stable for 12 months. RNase A is supplied lyophilized. On addition of Resuspension Buffer P1, the RNase A solution should be kept stored at 4°C, and is stable at this temperature for at least 6 months.