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cloning

Restriction Endonucleases

Description:
Invitrogen’s restriction endonucleases are rigorously tested to ensure quality performance. During production, the following four assays are performed:
  • Unit determination assay
  • Endonuclease assay
  • Exonuclease assay
  • Ligation-recut assay
The table below lists enzymes currently available.

Contents and Storage:
Each restriction endonuclease is supplied with the recommend 10X REact® buffer. Store at -20°C.
Enzyme Cut Site Reaction Conditions Reaction Conditions Heat inactivated* Methylation sensitive†
Acc I 5´-GT ↓ (A/C )(T/G ) AC-3´
3´-CA (T/G )(A/C) ↑ TG-5´
 REact® 1 Buffer at 37°C REact® 1 Buffer  at 37°C N Y
Alu I 5´-AG ↓ CT-3´
3´-TC ↑ GA-5´
 REact® 1 Buffer at 37°C REact® 1 Buffer  at 37°C 10 min.@ 65°C Y
Apa I 5´-G GGCC ↓ C-3´
3´-C ↑ CCGG G-5´
 REact® 4 Buffer at 30°C REact® 4 Buffer  at 30°C 10 min.@ 65°C Y
Ava I 5´-C ↓ PyCGPu G-3´
3´-G PuGCPy ↑ C-5´ 		REact® 2 Buffer at 37°C REact® 2 Buffer at 37°C 10 min.@ 65°C Y
BamH I 5´-G ↓ GATC C-3´
3´-C CTAG ↑ G-5´
 REact® 3 Buffer at 37°C REact® 3 Buffer  at 37°C N Y
Bgl II 5´-A ↓ GATC T-3´
3´-T CTAG ↑ A-5´ 		REact® 3 Buffer at 37°C REact® 3 Buffer  at 37°C N Y
Cla I 5´-AT ↓ CG AT-3´
3´-TA GC ↑ TA-5´
 REact® 1 Buffer at 37°C REact® 1 Buffer  at 37°C 10 min.@ 65°C dam, dcm
Dde I 5´-AT ↓ CG AT-3´
3´-TA GC ↑ TA-5´
 REact® 3 Buffer at 37°C REact® 3 Buffer at 37°C N Y
Dpn I 5´-GmA ↓ TC-3´
3´-C T ↑ mAG-5´
 REact® 4 Buffer at 37°C REact® 4 Buffer at 37°C 15 min.@ 75°C Y
Dra I 5´-TTT ↓ AAA-3´
3´-AAA ↑ TTT-5´
 REact® 1 Buffer at 37°C REact® 1 Buffer at 37°C Y N
EcoR I 5´-G ↓ AATT C-3´
3´-C TTAA ↑ G-5´
 REact® 3 Buffer at 37°C REact® 3 Buffer at 37°C 10 min.@ 65°C Y
EcoR V 5´-GAT ↓ ATC-3´
3´-CTA ↑ TAG-5´
 REact® 2 Buffer at 37°C REact® 2 Buffer at 37°C 10 min.@ 65°C Y
Hae III 5´-GG ↓ CC-3´
3´-CC ↑ GG-5´
 REact® 2 Buffer at 37°C REact® 2 Buffer at 37°C N Y
Hha I 5´-G CG ↓ C-3´
3´-C ↑ GC G-5´
 REact® 2 Buffer at 37°C REact® 2 Buffer at 37°C 10 min.@ 65°C Y
Hinc II 5´-GTPy ↓ PuAC-3´
3´-CAPu ↑ PyTG-5´
 REact® 4 Buffer at 37°C REact® 4 Buffer at 37°C 10 min.@ 65°C Y
Hind III 5´-A ↓ AGCT T-3´
3´-T TCGA ↑ A-5´ 		REact® 2 Buffer at 37°C REact® 2 Buffer at 37°C N Y
Hinf I 5´-G ↓ ANT C-3´
3´-C TNA ↑ G-5´
 REact® 2 Buffer at 37°C REact® 2 Buffer at 37°C N Y
Hpa I 5´-GTT ↓ AAC-3´
3´-CAA ↑ TTG-5´
 REact® 4 Buffer at 37°C REact® 4 Buffer at 37°C N Y
Hpa II 5´-C ↓ CG G-3´
3´-G GC ↑ C-5´
 REact® 8 Buffer at 37°C REact® 8 Buffer at 37°C N Y
Kpn I 5´-G GTAC ↓ C-3´
3´-C ↑ CATG G-5´
 REact® 4 Buffer at 37°C REact® 4 Buffer at 37°C 10 min.@ 65°C Y
Mbo I 5´- ↓ GATC -3´
3´- CTAG ↑ -5´
 REact® 2 Buffer at 37°C REact® 2 Buffer at 37°C 10 min.@ 65°C dam, dcm
Mlu I 5´- A ↓ CGCG T-3´
3´- T GCGC ↑ A-5´
 REact® 3 Buffer at 37°C REact® 3 Buffer at 37°C 10 min.@ 65°C Y
Msc I 5´-TGG ↓ CCA-3´
3´-ACC ↑ GGT-5´
 REact® 2 Buffer at 37°C REact® 2 Buffer at 37°C 10 min.@ 65°C dam, dcm
Mse I 5´-T ↓TA A-3´
3´-A AT ↑T-5´
 REact® 1 Buffer at 37°C REact® 1 Buffer at 37°C 10 min.@ 65°C N
Nco I 5´-C ↓CATG G-3´
3´-G GTAC ↑C-5´
 REact® 3 Buffer at 37°C REact® 3 Buffer at 37°C 10 min.@ 65°C Y
Nde I 5´-CA ↓TA TG-3´
3´-GT AT ↑AC -5´
 REact® 6 Buffer at 37°C REact® 6 Buffer at 37°C N N
Nhe I 5´-G ↓CTAG C-3´
3´-C GATC ↑G-5´
 REact® 4 Buffer at 37°C REact® 4 Buffer at 37°C N U
Not I 5´-GC ↓GGCC GC-3´
3´-CG CCGG ↑CG-5´
 REact® 3 Buffer at 37°C REact® 3 Buffer at 37°C 10 min.@ 65°C Y
Nru I 5´-TCG ↓ CGA-3´
3´-AGC ↑ GCT-5´
 REact® 7 Buffer at 37°C REact® 7 Buffer at 37°C 10 min.@ 65°C Y
Nsi I 5´-A TGCA ↓ T-3´
3´-T ↑ ACGT A-5´
 REact® 3 Buffer at 37°C REact® 3 Buffer at 37°C N N
PinA I 5´-A ↓ CCGG T -3´
3´-T GGCC ↑ A -5´
 REact® 4 Buffer at 37°C REact® 4 Buffer at 37°C 10 min.@ 65°C N
Pst I 5´-C TGCA ↓ G-3´
3´-G ↑ ACGT C-5´
 REact® 2 Buffer at 37°C REact® 2 Buffer at 37°C 10 min.@ 65°C Y
Pvu I 5´-CG AT ↓ CG-3´
3´-GC ↑ TA GC-5´
 REact® 7 Buffer at 37°C REact® 7 Buffer at 37°C N Y
Pvu II 5´-CAG ↓ CTG-3´
3´-GTC ↑ GAC-5´
 REact® 6 Buffer at 37°C REact® 6 Buffer at 37°C 10 min.@ 65°C Y
Rsa I 5´-GT ↓ AC -3´
3´-CA ↑ TG -5´
 REact® 1 Buffer at 37°C REact® 1 Buffer at 37°C 10 min.@ 65°C N
Sal I 5´-G ↓ TCGA C-3´
3´-C AGCT ↑ G-5´
 REact® 10 Buffer at 37°C REact® 10 Buffer at 37°C N Y
Sca I 5´-AGT ↓ ACT-3´
3´-TCA ↑ TGA-5´
 REact® 6 Buffer at 37°C REact® 6 Buffer at 37°C 10 min.@ 65°C N
Sma I 5´-CCC ↓ GGG-3´
3´-GGG ↑ CCC-5´
 REact® 4 Buffer at 30°C REact® 4 Buffer 10 min.@ 65°C Y
Spe I 5´-A ↓ CTAG T-3´
3´-T GATC ↑ A-5´
 REact® 4 Buffer at 37°C REact® 4 Buffer at 37°C 10 min.@ 65°C N
Sph I 5´-G CATG ↓ C-3´
3´-C ↑ GTAC G-5´
 REact® 6 Buffer at 37°C REact® 6 Buffer at 37°C 10 min.@ 65°C Y
Ssp I 5´-AAT ↓ ATT-3´
3´-TTA ↑ TAA-5´
 REact® 6 Buffer at 37°C REact® 6 Buffer at 37°C 10 min.@ 65°C N
Sst I 5´-G AGCT ↓ C-3´
3´-C ↑ TCGA G-5´
 REact® 2 Buffer at 37°C REact® 2 Buffer at 37°C 10 min.@ 65°C Y
Sst II 5´-CC GC ↓ GG-3´
3´-GG ↑ CG CC-5´
 REact® 2 Buffer at 37°C REact® 2 Buffer at 37°C N N
Stu I 5´-AGG ↓ CCT-3´
3´-TCC ↑ GGA-5´
 REact® 2 Buffer at 37°C REact® 2 Buffer at 37°C 10 min.@ 65°C dam, dcm
Taq I 5´-T ↓ CG A-3´
3´-A GC ↑ T-5´
 REact® 2 Buffer  at 65°C REact® 2 Buffer  at 65°C N Y
Xba I 5´-T ↓ CTAG A-3´
3´-A GATC ↑ T-5´
 REact® 2 Buffer at 37°C REact® 2 Buffer at 37°C N Y
Xho I 5´-C ↓ TCGA G-3´
3´-G AGCT ↑ C-5´
 REact® 2 Buffer at 37°C REact® 2 Buffer at 37°C N Y

REact® buffers

  • The appropriate REact® buffer is supplied with every Invitrogen restriction enzyme. The buffer compositions shown below are at 1X concentration.  

    REact®  1:    50 mM Tris-HCl (pH 8.0)
    10 mM MgCl2

    REact®  2:    50 mM Tris-HCl (pH 8.0)
    10 mM MgCl2
    50 mM NaCl

    REact® 3:    50 mM Tris-HCl (pH 8.0)
    10 mM MgCl2
    100 mM NaCl

    REact® 4:    20 mM Tris-HCl (pH 7.4)
      5 mM MgCl2
    50 mM KCl

    REact® 5:    10 mM Tris-HCl (pH 8.2)
      8 mM MgCl2

    REact® 6:    50 mM Tris-HCl (pH 7.4)
      6 mM MgCl2
    50 mM NaCl
    50 mM KCl

    REact® 7:     50 mM Tris-HCl (pH 8.0)
    10 mM MgCl2
    50 mM NaCl
    50 mM KCl

    REact® 8:     20 mM Tris-HCl (pH 7.4)
    10 mM MgCl2

    REact® 9:     20 mM TrisOAc (pH 7.9)
    10 mM MgOAc
    50 mM KOAc

    REact® 10:    100 mM Tris-HCl (pH 7.6)
    10 mM MgCl2
    150 mM NaCl

    REact® 11:    10 mM Tris-HCl (pH 9.0)
    12 mM MgCl2
    100 mM KCl