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Restriction enzyme noti7/31/2023 NOT FOR USE IN DIAGNOSTIC PROCEDURES (EXCEPT AS SPECIFICALLY NOTED). Our mission is to develop high-quality innovative tools and services to accelerate discovery.įOR RESEARCH USE ONLY. As a member of the Takara Bio Group, Takara Bio USA is part of a company that holds a leadership position in the global market and is committed to improving the human condition through biotechnology. provides kits, reagents, instruments, and services that help researchers explore questions about gene discovery, regulation, and function. Compositions of provided universal buffers and other reagents Since BSA and Triton X-100 are supplied with enzymes at a 10X concentration (0.1%), be sure to add 1/10 the volume of the reaction mixture to the buffer before starting the reaction. Some restriction enzymes are supposed to exhibit 100% activity when BSA or Triton X-100 is added to the reaction system. Since 10X T4 buffer does not contain BSA, be sure to add BSA to a final concentration of 0.01%. Please add 1/10 the volume of the reaction mixture. Universal buffers are provided at a 10X concentration. Instructions for the use of universal buffers If there is a precipitate, dissolve in a warm water bath before use. Note: SDS may precipitate during storage at room temperature. The tables below list relative activities of restriction enzymes in the universal buffers and instructions for use and compositions of buffers.ġ/10 volume of 10X loading buffer to stop the digestion reaction, and run on an agarose gel. The compositions of these basal buffers vary depending on the enzyme.įor information on double digestion, see the universal buffers for double digestion with restriction enzymes page. In order to avoid these effects, use of buffers highlighted in blue or pink is recommended.Ī few specific enzymes (AccIII, BalI, BcnI, BglI, Bpu1102I, Cfr10I, Eco52I, NruI, PshBI, SnaBI, SspI, TaqI, and VpaK11BI), are each supplied with a basal buffer specialized for the particular enzyme. Values in ( ) indicate buffers that are likely to be affected by star activity. Restriction Enzyme Digest Problem: Too Many DNA Bands Double Digestion with NEBcloner Reduce Star Activity with High-Fidelity Restriction Enzymes Standard Protocol for Restriction Enzyme Digests NEB ® TV Ep. The relative activity in each of the other universal buffers is normalized to the optimal buffer, where the activity of each enzyme in the optimal buffer is expressed as 100%. concentration measurements in UV light) the colorless FastDigest Buffer is recommended.įor methylation sensitivity, refer to product specifications.Our restriction enzymes are supplied with an optimal universal buffer (one of five universal buffers indicated in blue in the table below). For applications that require product analysis by fluorescence excitation (e.g. Note: The FastDigest Green Buffer offers the same high performance in DNA digestion and downstream applications as the colorless FastDigest Buffer. Restriction fragment length polymorphism (RFLP).100% buffer compatibility with downstream applications.100% activity of all FastDigest enzymes in the universal buffer.Short incubation times and optimal composition of the universal FastDigest Buffer eliminate star activity effects. Therefore, enzymes for downstream applications can be directly added to the FastDigest reaction mix.įor additional convenience FastDigest Green Buffer includes a density reagent and two tracking dyes for direct loading of digestion reaction products on gels. DNA modifying enzymes, such as Klenow Fragment, T4 DNA Ligase, alkaline phosphatases and T4 DNA Polymerase all have 100% activity in FastDigest Buffer. See Reaction Conditions for FastDigest Enzymes for a table of enzyme activity, heat inactivation and incubation times for this and other FastDigest restriction enzymes. The universal buffer allows rapid single-, double-, or multiple DNA digestion within 5–15 minutes eliminating any need for buffer change or subsequent DNA clean-up steps. Thermo Scientific FastDigest NotI is one of an advanced line of fast restriction enzymes that are all 100% active in the universal FastDigest and FastDigest Green reaction buffers. Thermo Scientific FastDigest NotI restriction enzyme recognizes GC^GGCCGC site and cuts best at 37☌ in 5–15 minutes using universal FastDigest Buffer.
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