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OPTIZYME™ SmaI, Fisher BioReagents™


Manufacturer: fisher scientific company  BP8011-5

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Catalog No. BP8011-5

  • / Each
 

Description & Specifications

Specifications

Product Name SmaI
Format Liquid
Concentration 10U/μL
Cut Site CCC.GGG
Incubator Temperature 30°C
pH 7.4
Includes 10X OPTIZYME™ Buffer 4
Quantity 5000U
Units 5,000U
For Use With (Application) >90% of pUC19 DNA fragments can be ligated and re-cut after 50-fold over-digestion with SmaI
Storage Buffer 10mM Tris HCl (pH 7.4 at 25°C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/mL BSA, 50% Glycerol

5'...C C C^G G G...3'
3'...G G G^C C C...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 30°C
Enzyme Activity in OPTIZYME buffers:
  • Buffer 1: 50 - 100%
  • Buffer 2: 0 - 20%
  • Buffer 3: 0 - 20%
  • Buffer 4: 100%
  • Buffer 5: 0 - 20%
Storage Buffer:
  • 10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA and 50% (v/v) glycerol
  • Ligation and Re-cleavage:
  • More than 90% of the pUC19 DNA fragments can be ligated and re-cut after 50-fold over-digestion with SmaI.
  • Digestion of Agarose-embedded DNA:
  • A minimum of 5 units of SmaI is required for the complete digestion of 1μg of agarose-embedded lambda DNA in 16 hours.
  • Notes:
    • Incubation at 37°C results in 50% activity. SmaI has a half-life of 15 min at 37°C.
    • Incubation at 25°C results in 100% activity.
    • SmaI needs K+ to work for activity.
    Methylation Effects:
    Dam: Never overlaps - no effect
    Dcm: Never overlaps - no effect
    CpG: Completely overlaps - blocked
    • CCCGGG
  • (Cleavage blocked)
  • EcoKI: Never overlaps - no effect
  • EcoBI: Never overlaps - no effect