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Support >  Applications >  Restriction_Enzymes >  Ligation-Recutting_Efficiency

Ligation Recutting Efficiency

Ligation-Recutting Efficiency

To determine the functional purity of an enzyme, a ligation-recutting assay was conducted. As shown in the chart below, the high quality of Takara's restriction enzymes are guaranteed by the criteria.

Contamination of ligase inhibitor, phosphatase and exonuclease was detected by the following procedures:

At first, overdigestion was performed by applying 2-fold to 50-fold excessive amount of restriction enzymes to a substrate DNA. Digested DNA was recovered and diluted to T4 DNA Ligase Buffer [66 mM Tris-HCl (pH7.6), 6.6 mM MgCl2, 10 mM DTT, 0.4 mM ATP] at a 5'-termini concentration of 0.1-1.0 µM. After an adequate amount of T4 DNA ligase was added, it was incubated at 16°C for 1 hour or 16-18 hours. The recovered DNA was diluted to a reaction buffer again and recut by the same restriction enzyme.

Restriction Enzyme Ligation Efficiency (%) Recutting Efficiency (%)
Aat II >90 >95
Acc I >90 100
Acc II >90 100
Acc III >95 100
Afa I 90 100
Afl II >95 100
Alu I >95 100
Aor13H I >95 100
Aor51H I 90 95
Apa I >95 100
ApaL I >95 100
Ava I >90 100
Ava II >90 100
Bal I >90 100
BamH I >95 100
Ban II 100 100
Bcn I* 90 90
Bgl I >95 100
Bgl II >95 100
Bln I 90 100
Bpu1102 I 90 100
BspT104 I >95 100
BspT107 I >95 100
Bsp1286 I 90 100
Bsp1407 I 90 100
BssH II 90 100
BstP I >95 100
BstX I >90 100
Bst1107 I 90 90
Cfr10 I 90 100
Cfr13 I 90 100
Cla I >95 100
Cpo I 100 100
Restriction Enzyme Ligation Efficiency (%) Recutting Efficiency (%)
Dra I >90 100
Eae I 100 100
Eam1105 I 90 90
EcoO65 I 95 100
EcoO109 I >95 100
EcoR I 100 100
EcoR V 90 100
EcoT14 I 100 100
EcoT22 I 90 100
Eco52 I 90 100
Eco81 I* 90 90
Fba I 100 100
Fok I >95 100
Fse I >95 100
Hae II >95 100
Hae III >95 100
Hap II >90 100
Hha I >90 100
Hinc II >95 100
Hind III 100 100
Hinf I >90 100
Hinl I 90 100
Hpa I >95 100
Kpn I >90 >95
Mbo I 95 100
Mbo II 90 100
Mfl I >95 100
Mlu I 100 100
Msp I 90 100
Mun I 90 100
Mva I* 90 90
Nae I 90 100
Nco I >95 100
Restriction Enzyme Ligation Efficiency (%) Recutting Efficiency (%)
Nde I 90 100
Nhe I 90 95
Not I >95 100
Nru I 95 95
Nsb I >90 >90
PmaCI 95 100
PshA I >90 100
PshB I 95 100
Psp1406 I 90 100
Pst I >95 100
Pvu I >95 100
Pvu II >95 100
Sac I >99 100
Sac II >90 95
Sal I >95 100
Sau3A I >95 100
Sca I >95 100
Sfi I 90 100
Sma I >95 100
SnaB I 90 >95
Spe I >99 100
Sph I 100 100
Sse8387 I 100 100
Ssp I 90 100
Stu I >95 100
Swa I >95 >95
Taq I >90 >90
Tth111 I >90 >90
Van91 I 90 100
VpaK11BI >90 100
Xba I 100 100
Xho I >95 100
Xsp I >90 100

*Digested DNA was ligated by TaKaRa DNA Ligation Kit, Ver.2 and incubated at 26°C for 10 min.

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