Topoisomerase I (E. coli)

Catalog # Concentration Size List Price Quantity Your Price
M0301L 5000 units/ml 500 units $477.00
$429.30
M0301S 5000 units/ml 100 units $117.00
$105.30
Catalog # Size List Price Your Price
M0301L 500 units $477.00
$429.30
M0301S 100 units $117.00
$105.30
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca
  • Catalyzes the relaxation of negatively-supercoiled DNA
  • Isolated from a recombinant source
  • Supplied with 10X Reaction Buffer
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Topoisomerase I (E. coli) catalyzes the relaxation of negatively supercoiled DNA. Topoisomerase I has also been implicated in knotting and unknotting DNA (1) and in linking complementary rings of single-stranded DNA into double-stranded rings (2). The intact holoenzyme is a 97 kDa protein.
Product Source
An E. coli strain containing the cloned topA gene
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  Topoisomerase I (E. coli) M0301SVIAL -20 1 x 0.02 ml 5,000 units/ml
  rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X
  Topoisomerase I (E. coli) M0301LVIAL -20 1 x 0.1 ml 5,000 units/ml
  rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme that catalyzes the relaxation of  > 95% of 0.5 μg of pUC19 RF I (negatively supercoiled) DNA in 15 minutes at 37°C in a total reaction volume of 25 μl. DNA supercoiling is assessed by agarose gel electrophoresis in the absence of ethidium bromide.

Reaction Conditions

1X rCutSmart™ Buffer
Incubate at 37°C

1X rCutSmart™ Buffer
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
100 µg/ml Recombinant Albumin
(pH 7.9 @ 25°C)

Activity in NEBuffers

NEBuffer™ 1: 25%
NEBuffer™ 2: 50%
NEBuffer™ 3: 0%
NEBuffer™ 4: 100%

Storage Buffer

10 mM Tris-HCl
35 mM (NH4)2SO4
50 mM KCl
1 mM DTT
0.1 mM EDTA
50% Glycerol
pH 7.5 @ 25°C

Heat Inactivation

65°C for 20 minutes

References
  • Liu, L.F., Depew, R.E. and Wang, J.C. (1976). J. Mol. Biol.. 106, 439-452.
  • Kirkegaard, K. and Wang, J.C. (1978). Nucl. Acids Res.. 5, 3811-3820.
Quality Control Assay
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
Specifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
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This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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