5´-GC↓GGCCGC-3´ 3´-CGCCGG↑CG-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	Not I 10X Buffer H (1mL) 10X Loading Buffer (1mL)
Reaction Temperature	37°C
Reaction Mixture	Not I 1 µL 10X Buffer H 2 µL 0.10% BSA 2 µL 0.10% Trion X-100 2 µL Substrate DNA <1 µg Sterile water to 20 µL
Heat inactivation	Enzyme is inactivated by heating at 65°C for 20 minutes.
Star Activity	Unrelated site may be under conditions of low ionic strength, high enzyme concentration, glycerol concentration >5%, or pH >8.0.
Effect of DNA Methylation	Enzyme activity is affected by CG methylase depending on the sequence of the recognition site or the sequence following the site.
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	Nocardia otididis-caviarum

Relative Activity in Universal Reaction Buffers

Buffer	L	M	H	K	T(+BSA)
Relative Activity (%)	<20*	<20*	20**	<20	<20*
*Weak star activity is detected. **100% activity is obtained by addition of 0.01% BSA and 0.01% Triton® X-100.

For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use.