Topoisomerase I - FAQs

View additional product information for Topoisomerase I - FAQs (38042024)

3 product FAQs found

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.