Ultrapure 100 Column

For in-house purification of up to 100 mg endotoxin-free ultrapure plasmid DNA

Features and benefits

Endotoxins efficiently reduced to <0.1 EU/µg DNA
Up to 100 mg ultrapure DNA
DNA suitable for gene therapy research, transfection, and other sensitive applications
Quick, easy setup, and simple lab operation
No toxic or immunogenic reagents

Principle

The QIAGEN Ultrapure 100 Column (see figure "QIAGEN Ultrapure 100 Column") is designed for fast and efficient purification of up to 100 mg plasmid DNA. The system uses QIAGEN anion-exchange technology with integrated endotoxin removal to yield ultrapure DNA.

QIAGEN Ultrapure 100 Column

Procedure

The bacterial biomass is lysed under alkaline conditions and the lysate is cleared by centrifugation (see flowchart). Endotoxins are removed using the special Endotoxin Removal Buffer, and the lysate is loaded onto the Ultrapure 100 Column for DNA purification using QIAGEN Anion-Exchange Resin.

The Ultrapure 100 Buffer Set provides buffers that are certified to contain <10 EU/ml (as determined by the BioWhittaker Kinetic-QCL Test) for use with the column. The Ultrapure 100 Column is delivered with a detailed protocol, and direct technical support is provided on request. A centrifuge and a peristaltic pump capable of flow rates up to 20 ml/min or FPLC are needed to operate the system.

Applications

The Ultrapure 100 Column yields plasmid DNA of a purity at least equivalent to that obtained after 2x CsCl-gradient centrifugation (see table "Comparison of ultrapure plasmid preparation technologies", reference 2). The simple endotoxin-removal step ensures that the purified DNA contains <0.1 EU/µg DNA. The DNA is therefore ideal for transfection, gene therapy research (1), and other sensitive applications.

Comparison of ultrapure plasmid preparation technologies

	2x CsCl gradient centrifugation	QIAGEN Ultrapure 100 system

Yield	100 µg – 5 mg DNA	Up to 100 mg DNA
DNA quality	High	Equal or better than 2x CsCl
Endotoxin removal	Additional extraction steps required	Integrated into chromatographic process
Toxic chemicals	CsCl, ethidium bromide, phenol, chloroform	None
DNA-damaging agents	UV light, toxic reagents	None
Centrifuge required	High-speed ultracentrifuge	Conventional centrifuge

Cited References

1. Le Borgne, S. et al. (1998) In vivo induction of specific cytotoxic T lymphocytes in mice and Rhesus macaques immunized with a DNA vector encoding an HIV epitope fused with hepatitis B surface antigen. Virology 240, 304.

2. Schorr, J. et al. (1995) Plasmid DNA for human gene therapy and DNA vaccines. Annals New York Acad. Sci. 772, 271.

Ordering Information
Product	Contents	Cat. No.
Ultrapure 100 Column	Column (44 mm x 250 mm) containing QIAGEN Anion-Exchange Resin; for preparation of up to 100 mg plasmid DNA Note: Please add RMB6167.00 as Deposit for column(押金) when ordering 11100. The money will be returned when the column is sent back to QIAGEN.	11100 　 1009686

Ultrapure 500 (resin)	QIAGEN Anion-Exchange Resin for preparation of up to 500 mg plasmid DNA	11500

Ultrapure 100 Buffer Set	Buffers P1, P2, P3, QBT, QC, QN, ER, TE, Endotoxin-free water, RNase A; for preparation of up to 100 mg plasmid DNA	11910

Note:The Ultrapure 100 Column and Buffer Set are not available in all countries. A deposit is required on the column. QIAGEN cannot accept Ultrapure 100 Columns that have been used in L2–L4/S2–S4 containment facilities.