Takara DNA Topoisomerase I

Takara DNA Topoisomerase I

Applications

• Conversion and analysis of DNA conformation

Buffer Components

• 10X DNA Topoisomerase I Buffer: 350 mM Tris-HCl (pH 8.0), 720 mM KCl, 50 mM MgCl₂, 50 mM DTT, and 50 mM spermidine
• Additional Component: 0.1% BSA

Source

• Calf thymus

Storage

• –20°C

Unit Definition

One unit is the amount of the enzyme that completely relaxes 0.5 µg of supercoiled pBR322 DNA in 50 µl of the reaction mixture in 30 min at 37°C.

Concentration

• 20 units/µl

Notes

• The supplied reaction buffer includes spermidine at a final concentration of 5 mM. Without spermidine, enzyme activity may decrease to 1/20.
• When analyzing supercoiled DNA (e.g., pBR322 or φX174 RF I) treated with DNA Topoisomerase I, electrophoresis should be performed on agarose gel without EtBr, followed by EtBr staining. This prevents re-supercoiling and allows accurate activity measurement.
• White precipitate may form when adding 0.1% BSA directly to the buffer. Prepare the reaction mixture (final 0.01% BSA) in the following order:
  dH₂O → supplied buffer → 0.1% BSA → substrate DNA
• The enzyme tends to adhere to glass or plastic surfaces; therefore, BSA must be included in the reaction mixture.

Additional Product Information

Please refer to the product’s Certificate of Analysis for storage conditions, product components, and technical specifications.
Kit Components List and Certificates of Analysis are available under the Documents tab.

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FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.
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