Description: New England Biolabs provides a color-coded 10X NEBuffer with each restriction endonuclease to ensure optimal (100%) activity. Most of our enzymes are supplied with one of four standard NEBuffers. Occasionally, an enzyme has specific buffer requirements not met by one of the four standard NEBuffers, in which case the enzyme is supplied with its own unique NEBuffer. Some restriction endonucleases require BSA at a final concentration of 100 µg/ml for optimal activity. When required, BSA is supplied as a 10 mg/ml (100X) stock and should be added to the reaction mixture.
Restriction endonucleases that do not require BSA for optimal activity are not adversely affected if BSA is present in the reaction mixture.
Reagents Supplied: 10X NEBuffer I-SceI BSA (100X)
1X NEBuffer I-SceI:
10 mM Tris-HCl
10 mM MgCl2
1 mM Dithiothreitol pH 8.8 @ 25°C
16-Hour Incubation: A 50 µl reaction containing this reaction buffer at a 1X concentration and 1 µg of HaeIII digested ΦX174 RF I DNA incubated for 16 hours resulted in no detectable non-specific nuclease degradation.
Endonuclease Activity: Incubation of this reaction buffer at a 1X concentration with 1 µg ΦX174 RF I DNA for 4 hours at 37°C in 50 µl reactions resulted in less than 5% conversion to RF II.
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