OPTIZYME™ HindIII, Fisher BioReagents™

Description

5'...A^A G C T T...3'
3'...T T C G A^A...5'

Supplied with: 10X OPTIZYME Buffer 5
Conditions for 100% Activity:

• 1X OPTIZYME Buffer 5: 10mM Tris-HCl (pH 8.5 at 37°C), 10mM MgCl₂, 100mM KCl and 0.1mg/ml BSA
• Incubate at 37°C

Enzyme Activity in OPTIZYME buffers:

• Buffer 1: 0 - 20%
• Buffer 2: 20 - 50%
• Buffer 3: 0 - 20%
• Buffer 4: 50 - 100%
• Buffer 5: 100%

Storage Buffer:
10mM Tris-HCl (pH 7.5 at 25°C), 250mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA and 50% (v/v) glycerol
Ligation and Re-cleavage:
More than 95% of the DNA fragments can be ligated and recut after a 50-fold over-digestion with HindIII.
Digestion of Agarose-embedded DNA:
A minimum 5 units of HindIII is required for the complete digestion of 1μg of agarose-embedded lambda DNA in 16 hours.
Methylation Effects:
Dam: Never overlaps - no effect
Dcm: Never overlaps - no effect
CpG: Never overlaps - no effect
EcoKI: Never overlaps - no effect
EcoBI: May overlap - cleavage impaired

• 5'...TGm6AAGCTT(N)₃ TGCT...3'

3'...AC TTCGAA(N)₃m6ACGA...5'

(Cleavage is impaired)