Home Takara Clontech Logo Takara Clontech
login | register | order now | view cart | feedback
Support
Applications


Support >  Applications >  Restriction_Enzymes >  Buffer_Activity_Chart

Restriction Enzyme Activity Displaying System using Universal and Basal Buffers

Buffer Activity Chart: Relative Restriction Enzyme Activity in Universal and Basal Buffers

Takara supplies an optimal universal buffer with each restriction enzyme (indicated in blue below). The relative activity in each of the other universal buffers is normalized to the optimal buffer, where activity of each enzyme in the optimal buffer is expressed as 100%.

Values in ( ) indicate buffers which are likely to be affected by star activity etc. In order to avoid these effects, use of buffers highlighted in blue or pink is recommended. For these enzymes (Acc III, Bal I, Bcn I, Bgl I, Bpu1102 I, Cfr10 I, Eam1105 I, Eco52 I, Nru I, PshB I, SnaB I, Ssp I, Taq I or VpaK 11 BI), basal buffer specialized for each enzyme is supplied.

The compositions of basal buffers vary depending on enzymes. For reference to double digestion, recommendable enzymes for each buffer (relative activity for each reaction mixture enzyme [see blue and pink highlighted items]) are listed. For more information on double digestions, see the Double Digestion Buffers chart.

Relative Activity for each Reaction Buffer

blue indicates: buffer supplied with the restriction enzyme
pink indicates: alternative buffer recommended for use

Restriction enzyme Relative activities (%)
L M H K T+BSA Basal***
Aat II <20 <20 <20 <20 100 120
Acc I 20 100 <20 (<20) 160 80
Acc II (260) 100 <20 20 200 160
Acc III (<20) (<20) 20 (80) (<20) 100
Afa I 60 60 40 60 100 100
Afl II 20 80* <20 <20 140 120
Alu I 100 100 <20 40 200 120
Aor13H I <20 20 <20 80* 80 100
Aor51H I 80 100 <20 20 120 120
Apa I 100 <20 <20 <20 <20 120
ApaL I 100 20 <20 <20 120 120
Ava I (<20) 100 20 40 100 120
Ava II 80 100 <20 20 100 100
Bal I 20 20 <20 <20 40 100
BamH I (<20) <20 40 100 (<20) 80
Ban II (120) (120) 100 80 (100) 100
Bcn I <20 20 40 60 60 100
Bgl I <20 <20 20 40 <20 100
Bgl II <20 20 100 (100) (60) 100
Bln I <20 20 40 100 20 120
BmeT110 I <20 <20 20 100 <20 140
BmgT120 I <20 <20 100 40 <20 240
Bpu1102 I <20 <20 <20 40 60 100
BspT104 I 100 60 <20 <20 100 120
BspT107 I <20 20 80 100 20 100
Bsp1286 I 100 20 <20 <20 60 100
Bsp1407 I 20 60 20 20 100 100
BssH II 100 100 60 20 140 100
BstP I (<20) (60) 100 (100) (100) 100
BstX I <20 40 100 <20 <20 120
Bst1107 I (<20) 60 100 100 40 100
Cfr10 I (<20) (<20) (<20) 40 (20) 100
Cla I 40 100 120 100 60 100
Cpo I <20 <20 80 100 <20 100
Dra I 100 100 60 100 80 80
Eae I 60 100 <20 <20 120 160
Eam1105 I (<20) (40) 20 40 (40) 100
EcoO65 I (20) (60) 60* 40 40 100
EcoO109 I 100 60 <20 <20 100 160
EcoR I (20) (100) 100 (120) (80) 120
EcoR V (<20) (40) 100 (120) (40) 100
EcoT14 I (<20) (40) 100 120 (60) 100
EcoT22 I <20 20 100 (140) (20) 120
Eco52 I <20 <20 <20 <20 <20 100
Eco81 I <20 100 <20 <20 100 160
Fba I (<20) (<20) (80) 100 (20) 100
Fok I (20) (60) <20 <20 (200) 100
Hae II 80 100 <20 80 140 100
Hae III 60 100 100 60 100 100
Hap II 100 60 <20 <20 100 80
Hha I 80 100 100 120 120 100
Hinc II 20 100 20 40 100 80
Hind III (60) 100 <20 200 (100) 80
Hinf I 80 100 100 160 60 100
Hin1 I 40 80* <20 20 60 160
Hpa I <20 (40) 20 100 (80) 100
Kpn I 100 60 <20 <20 (100) 80
Mbo I 20 40 60 100 40 100
Mbo II 100 60 <20 <20 60 100
Mfl I 100 80 <20 <20 80 100
Mlu I 60 60 100 (100) 60 100
Msp I 80 80 <20 100 100 80
Mun I (200) 100* <20 <20 160 100
Mva I (<20) (40) 80 100 (20) 120
Nae I 100 <20 <20 <20 100 120
Nco I (40) (60) 20 60* (60) 160
Nde I <20 40 100 100 80 100
Nhe I (120) 100 <20 <20 (160) 100
Not I (<20) (<20) 20** <20 (<20) 100
Nru I 0 <20 20 20 <20 100
Nsb I 40 20 <20 60 100 100
PmaC I 100 80 <20 <20 100 100
PshA I 20 40 <20 100 60 160
PshB I (20) (40) 20 40 40 100
Psp1406 I 20 60 <20 <20 100 100
Pst I (<20) (60) 100 80 (20) 80
Pvu I (<20) (20) (40) 80* (40) 120
Pvu II (80) 100 40 <20 (40) 100
Sac I 100 60 <20 <20 80 80
Sac II 40 20 <20 <20 100 40
Sal I <20 <20 100 (20) <20 120
Sau 3A I (60) 80 100 <20 (80) 100
Sca I (<20) (<20) 100 (60) (<20) 100
Sfi I (40) 100 <20 <20 100 100
Sma I <20 <20 <20 <20 100 100
Smi I <10 <20 100 40 <10 100
SnaB I (20) (40) <20 <20 (40) 100
Spe I (80) 100 80 100 (80) 100
Sph I (20) (40) 100 120 (20) 100
Sse8387 I (120) 60* <20 <20 (60) 100
Ssp I (<20) (60) 40 (100) (80) 100
Stu I 60 100 60 80 140 100
Taq I 40 80 60 60 80 100
Tth111 I (20) 80 40 100 (80) 120
Van91 I <20 (20) 60 100 (60) 100
VpaK11B I <20 <20 60 (40) <20 100
Xba I <20 80* 20 <20 120 120
Xho I <20 60 100 160 60 100
Xsp I <20 60 <20 100 160 100

*+0.01% BSA→100% Afl II, EcoO65 I, Fok I, Hin1 I, Mun I, Nco I, Pvu I, Spl I, Sse8387 I, Xba I
**+0.01% BSA+0.01% Triton X-100→100% Not I
*** The composition of basal buffers varies depending on enzymes.

^ to top


Enzymes Classified By Universal Buffer

L M H K T+BSA
Afa I Acc I Ban II Afa I Aat II
Alu I Acc II Bgl II Aor13H I* Acc I
Aor51H I Afa I BmgT120 I BamH I Acc II
Apa I Afl II* BspT107 I Ban II Afa I
ApaL I Alu I BssH II Bcn I Afl II
Ava II Aor51H I BstP I Bln I Alu I
BspT104 I Ava I BstX I BmeT110 I Aor13H I
Bsp1286 I Ava II Bst1107 I BspT107 I Aor51H I
BssH II BspT104 I Cla I Bst1107 I ApaL I

Bsp1407 I Cpo I
Ava I
Dra I BssH II Dra I Cla I Ava II
Eae I Bst1107 I EcoO65 I* Cpo I
EcoO109 I
EcoR I Dra I Bcn I
Hae II Cla I EcoR V EcoT14 I Bpu1102 I
Hae III Dra I EcoT14 I Fba I BspT104 I
Hap II Eae I EcoT22 I Hae II Bsp1286 I
Hha I EcoO109 I Hae III Hae III Bsp1407 I
Hinf I Eco81 I Hha I Hha I BssH II
Kpn I Fok I* Hinf I Hind III
Mbo II
Mbo I Hinf I Cla I
Mfl I Hae II Mlu I Hpa I Dra I
Mlu I Hae III Mva I Mbo I Eae I
Msp I Hap II Nde I Msp I EcoO109 I
Nae I Hha I Not I** Mva I Eco81 I
PmaC I Hinc II Pst I Nco I*
Sac I Hind III Sal I Nde I Hae II



Nsb I
Stu I Hinf I Sau3A I Psh AI Hae III

Hin1 I* Sca I Pst I Hap II

Kpn I Smi I Pvu I* Hha I

Mbo II Spe I Spe I Hinc II

Mfl I Sph I Sph I Hinf I

Mlu I Stu I Stu I Hin1 I

Msp I Taq I Taq I Mbo II

Mun I* Van91 I Tth111 I Mfl I

Nhe I VpaK11B I Van91 I Mlu I

PmaC I Xho I
Xho I Msp I

Psp1406 I
Xsp I Mun I

Pvu II

Nae I

Sac I

Nde I




Nsb I

Sau3A I

PmaC I

Sfi I

Psp1406 I

Spe I

PshA I




Psp1406 I

Sse8387 I*

Sac I

Stu I

Sac II

Taq I

Sfi I

Tth111 I

Sma I

Xba I*

Stu I

Xho I

Taq I

Xsp I

Xba I




Xho I




Xsp I

*+0.01% BSA
**+0.01% BSA+0.01% Triton X-100

10× Loading Buffer

All Takara's restriction enzymes are supplied with 10X Loading Buffer(1 ml).

Composition


1%
SDS
50%
Glycerol
0.05%
Bromophenol Blue

Compositions of Universal Buffer

1. 10× L 100 mM Tris-HCl (pH7.5)
100 mM MgCl2
10 mM Dithiothreitol
2. 10× M 100 mM Tris-HCl (pH7.5)
100 mM MgCl2
10 mM Dithiothreitol
500 mM NaCl
3. 10× H 500 mM Tris-HCl (pH7.5)
100 mM MgCl2
10 mM Dithiothreitol
1,000 mM NaCl
4. 10× K 200 mM Tris-HCl (pH8.5)
100 mM MgCl2
10 mM Dithiothreitol
1,000 mM KCl
5. 10× T
(BSA-free)
330 mM Tris-acetate (pH7.9)
100 mM Mg-acetate
5 mM Dithiothreitol
660 mM K-acetate
6. 0.1% BSA solved in sterilized water
7. 0.1% Triton X-100

Instructions for use

Universal Buffers are 10X concentration. Please add 1/10 volume of the reaction mixture.
Since 10X T4 buffer does not contain BSA, be sure to add BSA to a final concentration of 0.01%.
Some restriction enzymes are supposed to exhibit 100% activity when BSA or Triton X-100 is added to the reaction system. Since BSA or Triton X-100 supplied with enzymes is 10X concentration (0.1%), be sure to add 1/10 volume of the reaction mixture like buffer before starting the reaction.

Instructions for use

Add > 1/10 volume of 10X Loading Buffer to stop enzyme reaction and apply on agarose gel electrophoresis. SDS may precipitate during the storage at room temperature. In case precipitates generate, dissolve in warm bath before use.

^ to top
Clontech is a Takara Bio Company © 2014 Privacy Policy | Terms & Conditions