Thermo Scientific FastDigest SfiI restriction enzyme recognizes GGCCNNNN^NGGCC site and cuts best at 50°C in 5–15 minutes using universal FastDigest Buffer.Thermo Scientific FastDigest SfiI is one of an advanced line of fast restriction enzymes that are all 100% active in the universal FastDigest and FastDigest Green reaction buffers.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. SeeReaction Conditions for FastDigest Enzymesfor a table of enzyme activity, heat inactivation and incubation times for this and other FastDigest restriction enzymes. DNA modifying enzymes, such asKlenow Fragment,T4 DNA Ligase, alkaline phosphatases andT4 DNA Polymeraseall have 100% activity in FastDigest Buffer. Therefore, enzymes for downstream applications can be directly added to the FastDigest reaction mix.For additional convenience FastDigest Green Buffer includes a density reagent and two tracking dyes for direct loading of digestion reaction products on gels.Short incubation times and optimal composition of the universal FastDigest Buffer eliminate star activity effects.Thermo Scientific FastDigest SfiI限制性内切酶可识别GGCCNNNN ^ NGGCC位点,并使用通用FastDigest缓冲液在50°C下5-15分钟内最佳切割。Thermo Scientific FastDigest SfiI是优异的快速限制酶系列之一,在通用FastDigest和FastDigest绿色反应缓冲液中均具有100%的活性。通用缓冲液允许5-15分钟内快速进行单,双或多个DNA消化,从而无需更换缓冲液或随后的DNA净化步骤。有关此酶和其他FastDigest限制酶的酶活性,热灭活和孵育时间的表格,请参见FastDigest酶的反应条件。 DNA修饰酶(例如Klenow片段,T4 DNA连接酶,碱性磷酸酶和T4 DNA聚合酶)在FastDigest Buffer中均具有100%的活性。因此,可将用于下游应用的酶直接添加到FastDigest反应混合物中。为了方便进一步使用,FastDigest 绿色缓冲液包括一种密度试剂和两种跟踪染料,可用于将消化反应产物直接加载到凝胶上。较短的孵育时间和通用FastDigest缓冲液的最佳组合消除了星号活性的影响。 |