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Alphabetic List of SBS Restriction Endonucleases
Restriction Endonucleases (C-M)
CspA I (Age I) ACCGGT
Cat. # Size Conc.
113-1 100 units 8-12u/µl
Source: Corynebacterium species
Buffer supplied: 10×CspA I
Substrate for unit definition: Lambda DNA
Reaction conditions: 10 mM Tris-HCl (pH 7.9, 25oC), 10 mM MgCl2,1 mM dithiothreitol,
1 mM dithiothreitol, 100 µg/ml BSA. Incubate at 37oC.
Storage buffer: 100 mM KCl, 10 mM Tris-HCl (pH 7.4, 25oC), 0.1 mM EDTA,
1 mM dithiothreitol, 200µg/ml BSA and 50% glycerol. Store at -20oC.
Absence of contaminants: Forty units of CspA I do not produce any unspecific cleavage
products after 16 hrs incubation with 1 µg of Lambda DNA at 37oC.
After ten-fold overdigestion with CspA I greater than 90% of
the DNA fragments can be ligated and recut with this enzyme.
Heat inactivation: 65oC for 20 minutes.
EcoR I GAATTC
Cat. # Size Conc.
114-1 5000 units 8-12u/µl
Source: E. coli RY 13
Buffer supplied: 10×EcoR I
Substrate for unit definition: Lambda DNA
Reaction conditions: 50 mM NaCl, 100 mM Tris-HCl (pH 7.4, 25oC), 5 mM MgCl2,
0.025% Triton X-100.. Incubate at 37oC.
Storage buffer: 300 mM NaCl, 5 mM KPO4 (pH 7.4), 0.1mM EDTA,
1 mM dithiothreitol, 0.15% Triton X-100, 200 µg/ml BSA and 50% glycerol. Store at -20oC.
Absence of contaminants: Ten units of EcoR I do not produce any unspecific cleavage
products after 16 hrs incubation with 1 µg of Lambda DNA at 37oC.
After 50-fold overdigestion with EcoR I, greater than 98% of
the DNA fragments can be ligated and recut with this enzyme.
Heat inactivation: 65oC for 20 minutes.
Star activity: Conditions of low ionic strength, high enzyme concentration,
glycerol concentration>5%, or pH>8.0 may result in star activity.
EcoR V GATATC
Cat. # Size Conc.
115-1 2,000 units 8-12u/µl
Source: Escherichia coli, J62plg 74
Buffer supplied: 10×M
Substrate for unit definition: Lambda DNA
Reaction conditions: 50 mM NaCl, 10 mM Tris-HCl (pH 7.9, 25oC), 10 mM MgCl2,
1 mM dithiothreitol, 100 µg/ml BSA. Incubate at 37oC.
Storage buffer: 50 mM KCl, 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA,
1 mM dithiothreitol, 200 µg/ml BSA and 50% glycerol. Store at -20oC.
Absence of contaminants: Fifty units of EcoR V do not produce any unspecific cleavage
products after 16 hrs incubation with 1 µg of Lambda DNA at 37oC.
After 20-fold overdigestion with EcoR V, greater than 95% of
the DNA fragments can be ligated and recut with this enzyme.
Heat inactivation: 80oC for 20 minutes.
Star activity: Conditions of low ionic strength, high enzyme concentration,
glycerol concentration>5%, or pH>8.0 may result in star activity.
Hind III AAGCTT
Cat. # Size Conc.
116-1 5000 units 8-12u/µl
Source: Haemophilus influenzae Rd
Buffer supplied: 10×M
Substrate for unit definition: Lambda DNA
Reaction conditions: 50 mM NaCl, 10 mM Tris-HCl (pH 7.9, 25oC), 10mM MgCl2,
1mM dithiothreitol. Incubate at 37oC.
Storage buffer: 50 mM KCl, 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA,
1 mM dithiothreitol, 500 µg/ml BSA and 50% glycerol. Store at -20oC.
Absence of contaminants: Fifty units of Hind III do not produce any unspecific cleavage
products after 16 hrs incubation with 1 µg of Lambda DNA at 37oC.
After 100-fold overdigestion with Hind III, greater than 98% of
the DNA fragments can be ligated and recut with this enzyme.
Heat inactivation: 65oC for 20 minutes.
Star activity: Star activity may be observed in the presence of Mn2+.
Hinf I GANTC
Cat. # Size Conc.
117-1 2,000 units 8-12u/µl
Source: An E. coli strain that carries the cloned Hinf I gene from Haemophilus influenzae Rf.
Buffer supplied: 10×H
Substrate for unit definition: Lambda DNA
Reaction conditions: 100 mM NaCl, 50 mM Tris-HCl (pH 7.9, 25oC) , 10 mM MgCl2,
1 mM dithiothreitol, 100 µg/ml bovine serum albumin and DNA. Incubate at 37oC.
Storage buffer: 50 mM KCl, 10mM Tris-HCl (PH 7.4, 25oC) , 0.1 mM EDTA,
1mM dithiothreitol, 200 µg/ml bovine serum albumin and 50 % glycerol. Store at -20oC
Absence of contaminants: Thirty units of Hinf I do not produce any unspecific cleavage
products after 16 hrs incubation with 1 µg of Lambda DNA at 37oC.
After ten-fold overdigestion with Hinf I, greater than 90% of
the DNA fragments can be ligated and recut with this enzyme.
Heat inactivation: 80oC for 20 minutes.
Hpa I GTTAAC
Cat. # Size Conc.
118-1 200 units 8-12u/µl
Source: An E.coli strain that carries the cloned Hpa I gene from Haemophilus parainfluenzae
Buffer supplied: 10×A
Substrate for unit definition: Lambda DNA
Reaction conditions: 50 mM potassium acetate, 20 mM Tris-acetate (pH 7.9, 25oC),
10 mM magnesium acetate, 1 mM dithiothreitol. Incubate at 37oC.
Storage buffer: 50 mM KCl, 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA,
1 mM dithiothreitol, 500 µg/ml BSA and 50% glycerol. Store at -20oC.
Absence of contaminants: Ten units of Hpa I do not produce any unspecific cleavage
products after 16 hrs incubation with 1 µg of Lambda DNA at 37oC.
After ten-fold overdigestion with Hpa I, greater than 95% of
the DNA fragments can be ligated and recut with this enzyme.
Heat inactivation: No.
Star activity: Conditions of high enzyme concentration or glycerol
concentration>5%, may result in star activity.
Kpn I GGTACC
Cat. # Size Conc.
119-1 2000 units 8-12u/µl
Source: Klebsiella pneumonia OK 8
Buffer supplied: 10×Kpn I
Substrate for unit definition: Lambda DNA (EcoR I digest)
Reaction conditions: 10 mM Tris-HCl (pH 7.0, 25oC), 10 mM MgCl2,
1 mM dithiothreitol, 0.01% Triton X-100, 100 µg/ml BSA. Incubate at 37oC.
Storage buffer: 50 mM KCl, 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA,
1 mM dithiothreitol, 200 µg/ml BSAand 50% glycerol. Store at -20oC.
Absence of contaminants: Ten units of Kpn I do not produce any unspecific cleavage
products after 16 hrs incubation with 1 µg of Lambda DNA at 37oC.
After ten-fold overdigestion with Kpn I, greater than 95% of
the DNA fragments can be ligated and recut with this enzyme.
Heat inactivation: No.
Star activity: Conditions of low ionic strength, high enzyme concentration,
glycerol concentration>5%, or pH>8.0 may result in star activity.
Mbo I (Sau3A I) GATC
Cat. # Size Conc.
120-1 200 units 8-12u/µl
Source: Streptomyces species
Buffer supplied: 110×Mbo I
Substrate for unit definition: Lambda DNA (dam-)
Reaction conditions: 100 mM KCl, 10 mM Tris-HCl (pH 8.0, 25oC), 10 mM MgCl2,
1mM dithiothreitol. Incubate at 37oC.
Storage buffer: 50 mM KCl, 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA,
1 mM dithiothreitol, 200 µg/ml BSA and 50% glycerol. Store at -20oC.
Absence of contaminants: Twenty units of Mbo I do not produce any unspecific cleavage
products after 16 hrs incubation with 1 µg of Lambda DNA at 37oC.
After ten-fold overdigestion with Mbo I, greater than 95% of
the DNA fragments can be ligated and recut with this enzyme.
Heat inactivation: 65oC for 20 minutes.
MspC I(Afl II) CTTAAG
Cat. # Size Conc.
121-1 500 units 8-12u/µl
Source: Micrococcus species
Buffer supplied: 10×SH
Substrate for unit definition: Lambda DNA (Hind III digest)
Reaction conditions: 150 mM NaCl, 10 mM Tris-HCl (pH 7.9, 25oC), 10 mM MgCl2,
1 mM dithiothreitol, 100 µg/ml BSA. Incubate at 37oC.
Storage buffer: 50 mM KCl, 10 mMoC Tris-HCl (pH 7.9, 25oC), 0.1 mM EDTA,
1 mM dithiothreitol, 200 µg/ml BSA and 50% glycerol. Store at -20oC.
Absence of contaminants: Fifty units of MspC I do not produce any unspecific cleavage
products after 16 hrs incubation with 1 µg of Lambda DNA (Hind III digest) at 37oC.
After 10-fold overdigestion with MspC I, greater than 95% of
the DNA fragments can be ligated and recut with this enzyme.
Heat inactivation: 65oC for 20 minutes.
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