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Invitrogen™ Topoisomerase I

Catalog No. 38042024
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500 U
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Catalog No. Quantity
38042024 500 U
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Catalog No. 38042024 Supplier Invitrogen™ Supplier No. 38042024
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Description

Catalyzes removal of superhelical turns from covalently closed DNA by a transient breakage and rejoining of phosphodiester bonds

  • Topoisomerase I is active in the presence of EDTA
  • Source: Purified from calf thymus
  • Performance and Quality Testing: Endodeoxyribonuclease, 3' and 5' exodeoxyribonuclease, and phosphatase assays; conversion of super-coiled DNA to relaxed DNA
  • Unit Definition: One unit catalyzes the conversion of 0.5μg of superhelical ΦX174 RF DNA to a relaxed state in 30 min. at 37°C
  • Unit Reaction Conditions: 50mM Tris-HCl (pH 7.5), 50mM KCl, 10mM MgCl2 , 0.1mM EDTA, 0.5mM DTT, 30 μg/mL BSA, 0.5μg ΦX174 RF DNA, and enzyme in 50μL for 30 min. at 37°C

Relaxing positively and negatively supercoiled DNA, Producing DNA topoisomers, Cloning, Restriction Enzyme Cloning

Order Info

Shipping Conditions: Wet ice

Specifications

Content And Storage Store in freezer (-5 to -30°C).
Shipping Condition Wet Ice
Enzyme Topoisomerase
Quantity 500 U
Product Type Topoisomerase I

Frequently Asked Questions (FAQs)

What are the applications and recommended protocols for Topoisomerase I?

Topoisomerase I has two general applications: to relax supercoiled DNA, and to generate samples with defined superhelical density. Generating samples with defined superhelical density is described in Analytical Biochemistry, v.122, p. 253, by Singleton and Wells (1982).

For relaxing supercoiled DNA, use the following protocol:

1. Add 0.5 µg supercoiled PhiX 174 DNA, 50 mM TrisHCl (pH 7.5), 50 mM KCl, 10 mM MgCl2, 0.5 mM DTT, 0.1 mM EDTA, 30 µg/mL BSA, and 1 unit Topoisomerase I to a final volume of 50 µL.
2. Incubate 30 mins at 37 degrees C.
3. Analyze the treated samples on a 1% agarose gel that does not contain ethidium bromide. Stain with ethidium bromide after electrophoresis is complete.
Do you offer a pre-made buffer for use with Topoisomerase I (Cat. No. 38042024)?

No, we do not offer a pre-made buffer for use with this product.

When editing a cycle on the Procise System, I can readily delete a step, but cannot seem to insert a function into the cycle. What am I doing wrong?

When you edit a flask cycle, be sure to highlight a flask function in order to insert it. Similarly, when editing a cartridge cycle, highlight a cartridge function in order to insert it.

For Research Use Only. Not for use in diagnostic procedures.