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 methodand 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 Mini Kit is tested on a lot-to-lot basis. The Kits are tested by isolation of plasmid DNA from 4 ml culture of E.coli DH5α transformed with the plasmid pBluescript (A600 >2 units/ml). More than 25 μg of plasmid DNA could be quantified with spectrophotometer. 1 µg of the purified plasmid is used on restriction enzyme digestion with Eco RI and digested DNA is checked by agarose gel analysis.

Kit Contents：Cat.No. / Kit Contents

APD100 (100 preps/kit)	APD300 (300 preps/kit)
PD1 Buffer : 25 ml PD2 Buffer : 25 ml PD3 Buffer : 45 ml W1 Buffer : 45 ml Wash Buffer(concentrated) : 25 ml Elution Buffer :6 ml RNase A (50mg/ml):50μg PD columns : 100pcs Collection Tubes : 100 pcs	PD1 Buffer : 65 ml PD2 Buffer : 75 ml PD3 Buffer : 100 ml W1 Buffer : 130 ml Wash Buffer(concentrated) : 50 ml Elution Buffer :30 ml RNase A (50mg/ml):130μg PD columns : 300pcs Collection Tubes : 300 pcs