Description: New England Biolabs supplies a 10X reaction buffer with all of its enzymes. At a 1X concentration this reaction buffer assures optimal activity of the enzyme.
1X NEBNext® dsDNA Fragmentase Reaction Buffer:
20 mM Tris-HCl
10 mM MgCl2
50 mM NaCl
0.15 % Triton X-100 pH 7.5 @ 25°C
Quality Control for Current Lot
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.
Protease Assay:
Incubation of at least 1X dsDNA Fragmentase buffer with 0.2 nmol of a standardized mixture of proteins, for 20 hours at 37°C, resulted in no proteolytic activity detected by SDS-PAGE.
Phosphatase Assay:
Incubation of 10 μl of at least 1X dsDNA Fragmentase Buffer in protein phosphatase assay buffer (1 M diethanolamine @ pH 9.8 and 0.5 mM MgCl2) containing 2.5 mM p-nitrophenyl phosphate at 37°C for 4 hours yields no detectable p-nitrophenylene anion as determined by spectrophotometric analysis at 405 nm.
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