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Cat. No.: NTBN-2k (for 2KU)

Recognition Site

GAGTCNNNNN
5'...G A G T C N N N N N↓...3'
3'...C T C A G N N N N N ...5'

Description

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Nt.BstNBI is a type of nicking endonuclease that cleaves only one strand of the dsDNA substrate; it generates a nick on the dsDNA substrate without cleaving the dsDNA.

Activity Definition

One unit (U) of activity is defined as the amount of enzyme required to completely convert 1 µg of supercoiled pUC19 DNA into its open circular form in a 50 μl reaction system at 55°C for 1 hour.

Quality Control

• Extended Incubation Test: Incubate 10 U of Nt.BstNBI with supercoiled pUC19 DNA substrate at 55°C for 16 hours. Verify that the open circular DNA remains unchanged through agarose gel electrophoresis.
• RNase Residual Detection: Incubate 10 U of Nt.BstNBI with 500 ng of RNA at 37°C for 1 hour. Use agarose gel electrophoresis to confirm that over 90% of the RNA remains intact.