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OPTIZYME™ ClaI (Bsu15I), Fisher BioReagents™


Manufacturer: fisher scientific company  BP8024-5

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Catalog No. BP8024-5

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Description & Specifications

Specifications

Product Name ClaI (Bsu15I)
Format Liquid
Concentration 10U/μL
Cut Site AT.CGAT
Incubator Temperature 37°C
pH 7.4
Includes >50% Water, 25 to 50% Glycerin
Quantity 2500U
For Use With (Application) >95% of DNA fragments can be ligated and re-cut after 50-fold over-digestion with ClaI
Storage Buffer 10mM Tris HCl (pH 8 at 25°C), 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.5mg/mL BSA, 50% Glycerol
Storage Requirements Keep container tightly closed

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