Spin Columns

30 ug DNA per column

High-Speed Plasmid Mini Kit 

 

Sample: 1-4 ml of bacterial culture

Yield: up to 30 mg of plasmid/cosmid DNA

Format: spin column

Operation time: 20 min

Elution volume: 50-100 ml

Cat. No. S00101

Store at room temperature (15 - 25°C )

For research used only

Introduction

High-Speed Plasmid Mini Kit is designed for rapid isolation of plasmid or cosmid DNA from 1-4 ml of bacterial cultures. In the process, the modified alkaline lysis method (1) and RNase treatment are used to get cleared cell lysate with minimal genomic DNA and RNA contaminants. In the presence of a chaotropic salt, the plasmid DNA in the lysate binds to glass fiber matrix in the spin column (2). The contaminants are washed with an ethanol-contained wash buffer and finally, the purified plasmid DNA is eluted by low salt elution buffer or water. The protocol does not require DNA phenol extraction and alcohol precipitation. Typical yields are 10-20 mg for high-copy number plasmid or 0.5-5 mg for low-copy number plasmid. The entire procedure can be completed in 20 minutes and the purified plasmid DNA is ready for restriction digestion, ligation, PCR, and sequencing reaction.

Quality Control

The quality of High-Speed 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 that contains the plasmid pBluescript (A600 > 2 units/ ml). More than 20 mg of plasmid DNA could be quantified with spectrophotometer. 1 mg of the purified plasmid is used on restriction enzyme digestion with Eco RI and digested DNA is checked by agarose gel analysis.

Kit Contents

Name	S00101	S00101-S
PD1 Buffer*	25 ml	1 ml
PD2 Buffer**	25 ml	1 ml
PD3 Buffer	45 ml	1.5 ml
W 1 Buffer	45 ml	2 ml
Wash Buffer (concentrated)***	25 ml	1 ml
Elution Buffer (10mM Tris-HCl, pH 8.5 at 25°C)	6 ml	1 ml
RNase A (50 mg/ ml)	50 ml	-
PD column	100 pcs	4 pcs
2 ml Collection tube	100 pcs	4 pcs

*   Add provided RNase A to PD1 Buffer and store at 4°C.  

** If precipitates have formed in PD2 Buffer, warm the buffer in a 37°C waterbath   to dissolve   precipitates. 

*** Add 100 ml/ 4 ml ethanol (96-100%) to Wash Buffer prior to initial use. 

  

Caution

The component contains irritant agent. During operation, always wear a lab coat, disposable gloves, and protective goggles.

  

References

(1) Birnboim, H. C., and Doly, J. (1979) Nucleic Acids Res. 7, 1513.

(2) Vogelstein, B., and Gillespie, D. (1979) Proc. Natl. Acad. Sci. USA 76, 615.

   

Spin Columns