5´-G↓AATTC-3´ 3´-CTTAA↑G-5´

Important: 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 Temperature	37°C
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

Buffer	L	M	H	K	T(+BSA)
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.