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

*+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.

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Enzymes Classified By Universal Buffer

10× Loading Buffer

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

Composition

Compositions of Universal Buffer

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.