OPTIZYME™ ClaI (Bsu15I), Fisher BioReagents™

Description

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

Supplied with: 10X OPTIZYME Buffer 4
Conditions for 100% Activity:

• 1X OPTIZYME Buffer 4: 33mM Tris-acetate (pH 7.9 at 37°C), 10mM Mg-acetate, 66mM K-acetate and 0.1mg/ml BSA
• Incubate at 37°C

Enzyme Activity in OPTIZYME buffers:

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

Storage Buffer:

• 10mM Tris-HCl (pH 8.0 at 25°C), 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.5mg/ml BSA and 50% (v/v) glycero

l

Ligation and Re-cleavage:

More than 95% of the DNA fragments can be ligated and re-cut after 50-fold over-digestion with ClaI.

Digestion of Agarose-embedded DNA:

A minimum of 5 units of ClaI is required for the complete digestion of 1μg of agarose-embedded lambda DNA in 16 hours.

Note:

• ClaI is blocked by overlapping dam methylation. To avoid dam methylation, use a dam-, dcm- strain.

Compatible Ends: Bsp119I, Hin1I, Hin6I, HpaII, MaeII, MspI, NarI, Psp1406I, SsiI, TaqI, XmiI

Methylation Effects:

Dam: May overlap - blocked

• 5'...ATCGm6A TC...3'

3'...TAGC Tm6AG...5'

(Cleavage is blocked)

Dcm: Never overlaps - no effect

CpG: Completely overlaps - blocked

• 5'...ATCGAT...3'

(Cleavage is blocked)

EcoKI: Never overlaps - no effect

EcoBI: May overlap - effect not determined

See Product Certificates