Takara 双酶切buffer用表

Spe I 1
×
M
1
×
K
1
×
H
1
×
M
1
×
H
1
×
H
1
×
M
1
×
M
1
×
M
1
×
K
+B
S
A
1
×
H
1 ×
H
+B
SA
1
×
H
1
×
K
1
×
M
1
×
H
1
×
T
+B
S
A
-
1
×
H
1
×
M
1
×
H
Sph I 0.
5
×
K
1
×
K
1
×
H
1
×
H
1
×
H
1
×
H
2
×
T
1
×
K
0.
5
×
K
1
×
K
+B
S
A
1
×
H
1 ×
H
+B
SA
1
×
H
1
×
K
0.
5
×
K
1
×
H
0.
5
×
T
+B
S
A
1
×
H
-
2
×
T
1
×
H
Xba I 1
×
M
0.
5
×
K
2
×
T
1
×
M
1
×
M
2
×
T
1
×
M
1
×
M
1
×
M
1
×
M
+B
S
A
1
×
T
0.5
×K
+B
SA
1
×
M
0.
5
×
K
1
×
M
1.
5
×
T
1
×
T
+B
S
A
1
×
M
2
×
T
-
1
×
M
Xho I 1
×
M
1
×
K
1
×
H
1
×
H
1
×
H
1
×
H
1
×
M
1
×
M
1
×
M
1
×
K
+B
S
A
1
×
H
1 ×
H
+B
SA
1
×
H
1
×
K
1
×
M
1
×
H
1
×
T
+B
S
A
1
×
H
1
×
H
1
×
M
-
Note:
1) It is confirmed that 10 units of each enzyme completely digest 1 µg of DNA at 37°C in one hour in 50 µl reaction mixture.
2) The concentration of Glycerol should be less than 10% to minimize star activity.
3) DNA may not be digested completely, when recognition sequences of two enzymes are close each other, or when DNA takes high-structure conformation.
Takara applies a system in which enzyme activities measured in the optimal universal buffers (one of 5 buffers:) are displayed. The relative activity in other universal buffers is expressed as 100% of the optimal universal buffers.
( ) indicates buffers which are likely to be affected by star activity etc. In order to avoid these effects, use of or buffers is recommended. For 15 enzymes (Acc III, Bal I, Bcn I, Bgl I, Bpu1102 I, Cfr10 I,
Eam1105 I, Eco52 I, Nru I, Psh B I, Sna B I, Ssp I, Taq I or Vpa K 11 BI,basal buffer specializedfor each enzyme is supplied).
The compositions of basal buffers vary depending on enzymes. For reference to double digestion, recomendable enzymes for each buffer (relative activity for each reaction mixture and enzyme) are listed.
Relative activity for each reaction buffer
: supplied, activity measured buffer
: recommended buffer
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 Apa L 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 Bam H I (<20) <20 40 100 (<20) 80 Ban II (120) (120) 100 80 (100) 100 Bbe I <20 <20 <20 <20 <20 100 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 Bme T110 I <20 <20 20 100 <20 140 Bmg T120 I <20 <20 100 40 <20 240 Bpu1102 I <20 <20 <20 40 60 100 Bsp T104 I 100 60 <20 <20 100 120 Bsp T107 I <20 20 80 100 20 100 Bsp1286 I 100 20 <20 <20 60 100
*+0.01% BSA→100% Afl II, Eco O65 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.
*+0.01% BSA
**+0.01% BSA+0.01% Triton X-100
10× Loading Buffer
All Takara's restriction enzymes are supplied with 10× Loading Buffer(1 ml).
Composition
1% SDS
50% Glycerol
0.05% Bromophenol Blue
Compositions of universal buffer
Instructions for use
Universal Buffers are 10× concentration. Please add 1/10 volume of the reaction mixture.
Since 10× T buffer does not contain BSA, be sure to add BSA to a final concentration of 0.01%.
Some restriction enzymes are supposed to exhibit a 100% activity when BSA or Triton X-100 is added to the reaction system. Since BSA or Triton X-100 supplied with enzymes is 10× 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 10× 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.。