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  Restriction endonucleases

Sca I

ScaI is a restriction enzyme purified from Streptomyces caespitosus.

Reagents supplied: 10x ScaI buffer

Unit definition: One unit is the amount of enzyme required to digest 1 μg of Lambda DNA in 60 min at 37oC in a total reaction volume of 50 μl.

(SKUs: RE033S, RE033L, REc033S, REc033L)

50.00120.00
  • 10u/μl
  • 40-60u/μl
  • 3×3000 U
  • 3000 U

Product Details

ScaI is a restriction enzyme purified from Streptomyces caespitosus.

Reagents supplied: 10x ScaI buffer and 10x eQ buffer

Unit definition: One unit is the amount of enzyme required to digest 1 μg of Lambda DNA in 60 min at 37oC in a total reaction volume of 50 μl.

Digest Guidelines

  • Standard reaction conditions: In order to overcome DNA source variability, a ~10 fold overdigest is suggested:
10U Sca I1μl
10x ScaI buffer5μl
DNA substrate1μg
Sterile ultrapure waterUp to 50 μl
  • Speed Digest reaction: Incubate for 15 minutes at 37oC as in standard reaction in 1x ScaI buffer (100 mM KCl, 10 mM Bis-Tris-Propane (pH 7.0 @ 25°C), 10 mM MgCl2, 100 μg/ml BSA) or 5 units in 2x eQ buffer (40 mM Tris-acetate (pH 7.9 @ 25oC), 20 mM Mg-acetate, 100 mM K-acetate, 200 μg/ml BSA) .
  • Activity(%) in EnzyQuest buffers -Double digest tips
R1R2R3R4EQ
<1025-5010075-10075-100

Reactions were performed as for unit definition reaction. The above data could be used as guideline for double digest reactions. Our enzymes are also active and compatible in buffer systems from other vendors.

Heat inactivation: 80oC for 20 minutes

Methylation Sensitivity:

  • dam methylation: Not sensitive
  • dcm methylation: Not sensitive
  • CpG methylation: Not sensitive

Unit calculation assay conditions:

  • 1x ScaI buffer
  • Lambda DNA
  • Incubation at 37oC

Storage conditions:

  • 50 mM KCl,
  • 10 mM Tris-HCl (pH 7.4)
  • 0.1 mM EDTA
  • 1 mM dithiothreitol
  • 500 μg/ml BSA
  • 50% glycerol
  • Store at -20oC

Non-specific nuclease activity: 600 units of Sca I do not produce any unspecific cleavage products after 16 hrs incubation with 1 μg of Lambda DNA at 37oC.

Re-ligation and recut: After 100-fold overdigestion with Sca I, greater than 90% of the DNA fragments can be ligated and recut with this enzyme.

Star activity: Large excess of the enzyme may results in the appearance of star activity.

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