Invitrogen restriction enzymes now come with Universal Buffers L, M, H, K, or T (+BSA) instead of REact® Buffers 1-8. Functionally, the enzymes are the same and have been validated for the recommended buffer. Please contact our technical services department immediately if you have questions or concerns.
|Concentration||30-60 U/ µL|
|Contents||EcoR I 10X Buffer H (1mL) 10X Loading Buffer (1mL) |
|Reaction Mixture ||EcoR I 1 µL 10X Buffer H 2 µL Substrate DNA < 1 µg Sterile water to 20 µL |
|Heat inactivation||Enzyme inactivated by heating at 65°C for 20 minutes.|
|Star Activity||Unrelated site may be cut in the presence of high concentrations of glycerol or Mn2+, and at low ionic strength. Enzyme activity reduces to 70-80% by the addition of spermine (0.2mM), but star activity reduces to 30-50%.|
|Effect of DNA Methylation||The sequence GAATTCG is affected by CG methylase.|
|Unit Definition||One unit is the amount of enzyme required to completely digest 1 µg of λDNA in 50 µL of the reaction mixture in one hour at 37°C.|
|10X Buffer H|| 500 mM Tris-HCl, pH 7.5 100 mM MgCl2 10 mM Dithiothreitol 1,000 mM NaCl |
|Source||Escherichia coli RY 13|
Relative Activity in Universal Reaction Buffers
|Relative Activity (%)||20*||100*||100||120*||80*|
* weak star activity is detected For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use.