ATP biotech Inc.

Introduction

Format: Spin columns
Sample: 1~4 ml bacterial cultures
Operation: Centrifuge / vacuum manifold
Operation time: 20 minutes
Binding capacity: Up to 30 µg per Column
Expected yield: 25~35 µg for high-copy plasmid;5~15 µg per low-copy plasmid
Application: DNA Library Screening and Analysis; DNA sequencing; Transformation; PCR; Restriction Digestion

ATP™ Plasmid Mini Kit is designed for the rapid , easily-handling, and cost-effective isolation of plasmid or cosmid DNA from 1~4 ml of bacterial cultures. For large scale plasmid purification we recommend ion exchange based Fast Ion Plasmid Mini/Maxi Kits.

1. This procedure uses modified alkaline lysis method
and RNase treatment for creating cleared cell lysate with minimal genomic DNA and RNA contaminants.
2. Subsequently, the lysate is neutralized and adjusted to high-salt binding conditions adaptable to the adsorption of DNA in one step. In the presence of a chaotropic salt, the plasmid DNA in the lysate binds to the uniquely-designed glass fiber matrix in the spin column .
3. Whereas RNA, cellular proteins, and other unwanted impurities flow through the column and are easily and efficiently removed from reaction mixture . After a brief washing step with ethanol-contained wash buffer to remove endonucleases, salts and other contaminants .
4. The purified plasmid DNA is eluted by a low salt elution buffer or water. The entire procedure can be completed in 20 ninutes and the purified plasmid DNA is immediately ready for restriction digestion, ligation, PCR, and sequencing.

The procedure does not require DNA phenol extraction and alcohol precipitation.

Quality Control

The quality of ATP™ Plasmid Midi/Maxi Kits is tested on a lot-to-lot basis. The Kits are tested by isolation of plasmid DNA from 50ml/100ml culture of E.coli DH5α transformed with the plasmid pBluescript (A600 >2 units/ml). More than 120μg/400μg of plasmid DNA could be quantified with spectrophotometer. 1 µ g of the purified plasmid is used on restriction enzyme digestion with EcoRI and digested DNA is checked by agarose gel analysis.

Kit Contents：Cat.No. / Kit Contents

APM010 (10 preps/kit)	APD300 (300 preps/kit)
PM1 Buffer : 25 ml PM2 Buffer : 25 ml PM3 Buffer : 45 ml PEQ Buffer : 45 ml PW Buffer(concentrated) : 25 ml PEL Buffer:130 mll RNase A (50mg/ml):200μl Plasmid-Maxi Columns : 10pcs	PM1 Buffer : 275 ml PM2 Buffer : 275 ml PM3 Buffer : 275 ml PEQ Buffer : 275 ml PW Buffer(concentrated) : 790 ml PEL Buffer :350 ml RNase A (50mg/ml):550μl Plasmid-Maxi Columns : 25pcs