Plasmid Substrates

All our DNA substrates are available separately and have been prepared to a high standard for accurate quantification of results. They are guaranteed for use with all our topoisomerase assay kits, buffers and enzymes.

Click on the product headings below to view details and order online.

Supercoiled pBR322 DNA

Supercoiled plasmid pBR322 DNA is produced by the large-scale alkaline-lysis method ( Sambrook et al., 1989)¹from a high-copy derivative of pBR322 (Boros et al., 1984)².

Negatively-supercoiled pBR322 DNA is further treated so that the DNA is more supercoiled and has a narrow range of linking numbers making it an ideal substrate for relaxation reactions.

Positively-supercoiled plasmid pBR322 DNA is produced by treating relaxed pBR322 with reverse gyrase before final purification.

It is shipped on dry ice at a concentration of 1mg/ml in TE (10mM Tris-HCl (pH 7.5), 1mM EDTA). Store at 4^oC

Technical Documents

Cat No.	Product	Price
S5001	Supercoiled pBR322 plasmid50 µg	£118
S2502	Supercoiled pBR322 plasmid250 µg	£430
S5003	Supercoiled pBR322 plasmid500 µg	£830
S1004	Supercoiled pBR322 plasmid1 mg	£1,595
POS5001	Positively Supercoiled pBR322 plasmid50 µg	£250
POS2502	Positively Supercoiled pBR322 plasmid250 µg	£990

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Relaxed pBR322 DNA

Relaxed plasmid pBR322 DNA is produced by the large-scale alkaline-lysis method ( Sambrook et al., 1989)1 from a high-copy derivative of pBR322 (Boros et al., 1984)2 and the resulting supercoiled plasmid is relaxed using topoisomerase I. This is further purified and optimised for topoisomerase assays

It is shipped on dry ice at a concentration of 1mg/ml in TE (10mM Tris-HCl (pH 7.5), 1mM EDTA). Store at 4^oC

0.5 µg of relaxed pBR322 when incubated with 1 U of DNA gyrase in a reaction volume of 30 µl at 37^oC for 30 minutes in incubation buffer is completely converted to the supercoiled form.

Technical Documents

Cat No.	Product	Price
R5001	Relaxed pBR322 plasmid50 µg	£118
R2502	Relaxed pBR322 plasmid250 µg	£430
R5003	Relaxed pBR322 plasmid500µg	£830
R1004	Relaxed pBR322 plasmid1 mg	£1,595

Custom Quote

Linear pBR322 DNA

Linear plasmid pBR322 DNA is produced by the large-scale alkaline-lysis method ( Sambrook et al., 1989)1 from a high-copy derivative of pBR322 (Boros et al., 1984)².

It is linearised by incubation with EcoRI and further concentrated and purified to a final concentration of of 1 mg/ml in TE (10 mM Tris-HCl (pH 7.5), 1 mM EDTA). Store at 4^oC

It is shipped on dry ice.

Technical Documents

Cat No.	Product	Price
L0150	Linear pBR322150 µg	£100
L0300	Linear pBR322300 µg	£178
L0600	Linear pBR322600 µg	£356
L1000	Linear pBR3221 mg µg	£556

Linking Number Plasmid

Plasmid pBR322 is partially negatively supercoiled to give specific linking numbers so that each marker has a range of 5-6 linking numbers. The kit contains a set of markers from relaxed plasmid covering approximately 23 linking numbers. Please enquire about single markers.

Technical Documents

Cat No.	Product	Price	Quantity
LNM001	Linking Number markersSet of 10 (20 µl of each at 1 µg/µl) enough for 10 gels	£240

References

Sambrook, J., Fritsch, E.F. & Maniatis, T (1989). Molecular cloning: a laboratory manual,2nd ed., Cold Spring Harbor Press. Cold Spring Harbor, NY.
Boros, I., Pósfai, G. & Venetianer, P. (1984). High-copy number derivatives of the plasmid cloning vector pBR322. Gene 30, 257-260
Trask, D. K. & Muller, M. T. (1983) Biochemical characterization of topoisomerase I purified from avian erythrocytes. Nucleic Acids Res. 11, 2779-2800
Shapiro, T.A., Klein, V.A. and Englund, P.T. (1999) Isolation of kinetoplast DNA, in DNA Topoisomerase Protocols Vol.I (Bjornsti, M-A and Osheroff, N., eds.). Humana Press Inc., N. Jersey, pp. 61-68

品牌介绍

Inspiralis

Inspiralis在提供高质量拓扑异构酶，连接酶和解旋酶方面享誉全球，包括检测试剂盒，底物和筛选服务。

We are located in laboratories adjacent to the academic research centres of the John Innes Centre and the University of East Anglia on the Norwich Research Park. We maintain close links with the academic research group of Professor Maxwell, helping us to remain at the forefront of the topoisomerase field. The experience of our staff and the connections with the academic group allow us to provide a high level of support to ensure that researchers are able to get the best results using our products. The same scientists who produce our enzymes and substrates also provide the technical support.

Our aim is to provide pharmaceutical companies, and others involved in drug development, with the necessary tools to develop and screen novel anti-infective and anti-cancer compounds. In order to achieve this we will continue to expand the range of topoisomerase products and screening services we offer.

产品列表：

No.	品牌	货号	名称	规格
1	Inspiralis	7F001	7F11 Monoclonal	0.1ml
2	Inspiralis	PA001	Anti-Gyr A Polyclonal	0.1 ml
3	Inspiralis	HT001	Anti-human topo II polyclonal - 0.1mL	0.1 ml
4	Inspiralis	G1001	E.coli gyrase	100 U
5	Inspiralis	T4001	E. coli Topo IV	100 U
6	Inspiralis	SAG4001	S. aureus gyrase	100 U
7	Inspiralis	SAT4001	S. aureus Topo IV	100 U
8	Inspiralis	MTG001	M. tuberculosis Gyrase (HIS)	100 U
9	Inspiralis	MTT01	M.tuberculosis Topo I	100 U
10	Inspiralis	PAG1001	P. aeruginosa Gyrase	100 U
11	Inspiralis	PAT4001	P. aeruginosa Topo IV	100 U
12	Inspiralis	SPG1001	S. pneumoniae Gyrase	100 U
13	Inspiralis	SPT4001	S. pneumoniae Topo IV	100 U
14	Inspiralis	ABG1001	A. baumannii Gyrase	100 U
15	Inspiralis	ABT4001	A. baumannii Topo IV	100 U