SgeI

SgeI

S750393-50μl

50μL

-20°C储存

超低温冰袋运输

-20ºC保存，两年有效。

SgeI可切割在单链或双链DNA上含有5-甲基胞嘧啶的DNA靶标。SgeI限制性内切酶可识别m5CNNG(9/13)^位点，并于37℃下在其独特的缓冲液中的切割效果最佳。为确保一致的性能，该酶储存缓冲液中包含预混合BSA，其可增强酶的稳定性，并与DNA制剂中可能存在的污染物相结合。 注意事项 ： 内切酶使用时宜存放在冰盒内或冰浴上，使用完毕后宜立即放置于-20ºC保存。底物至少需要2个SgeI识别序列才能有效酶切。甲基化DNA完全酶切取决于SgeI的识别位点的数量，另外由于识别位点酶切产生的DNA产物会促进SgeI的非特异性酶切，因此，建议酶切时优化SgeI酶量用于酶切反应。不含核酸酶的超纯水推荐选购阿拉丁的ST876 Ultrapure Water (DNase/RNase-Free, Sterile)。如果发现预期的酶切位点不能切开，请确认是否存在甲基化干扰问题。同裂酶对于不同的甲基化修饰可能具有不同敏感性，遇到可能存在甲基化干扰问题时，可以尝试同裂酶。本产品仅限于专业人员的科学研究用，不得用于临床诊断或治疗，不得用于食品或药品，不得存放于普通住宅内。为了您的安全和健康，请穿实验服并戴一次性手套操作。 使用说明 ： 1.单酶切时可以参考如下反应体系，在冰浴上进行操作。 Reagent Plasmid DNA Ultrapure Water to 20µl 10XSgeⅠBuffer 2µl Substrate DNA xµl (up to0.5- 2µg) SgeI 0.2-1µl Total volume 20µl Incubate at 37℃ 1h 注：反应体系可以按比例放大或缩小。反应时间不建议超过1h。a.参考上表依次加入各种液体后，用移液器轻柔吸打或轻弹管壁以混匀(切勿涡旋混合)，然后瞬时离心以沉降液体至管底。b.37℃温育1h。酶切反应时优先推荐使用水浴，反应温度通常更加恒定一些。c.80℃温育20min即可使酶失活并停止反应(可选)。2.双或多酶切时可以在参考上表单酶切反应体系设置的基础上，参考如下原则设置反应体系。a.每种内切酶的用量为1μl，并根据需要适当扩大反应体系；b.所有内切酶的体积总和不得超过总反应体系的1/10；c.如果所用的几种内切酶的最适反应温度不同，应先从最适温度低的酶开始酶切，再添加最适温度较高的酶，在其最适反应温度下进行酶切反应。 SgeI is a restriction endonuclease that can cleave DNA targets containing 5-methylcytosine on either single-stranded or double-stranded DNA. It recognizes the m5CNNG(9/13) site and exhibits optimal digestion efficiency in its unique reaction buffer at 37℃. To ensure consistent performance, the enzyme storage buffer contains BSA, which enhances enzyme stability and binds to any potential contaminants in the DNA preparation.Enzyme activity assay Precautions ： The restriction enzyme should be kept on ice during use, and stored at -20℃ immediately after use.The SgeI requires at least 2 recognition sites in the substrate DNA for efficient cleavage.Complete digestion of methylated DNA by SgeI depends on the number of recognition sites in the DNA. Additionally, the products generated from the specific cleavage can promote non-specific cleavage by SgeI. Therefore, it is recommended to optimize the amount of SgeI for the digestion reaction.For ultrapure water without nuclease, we recommend the Pure™ Ultrapure Water (DNase/RNase-Free, Sterile) (, ST876).If the expected enzyme digestion site cannot be cut, please check for potential methylation interference.Isoschizomers may have different sensitivities to different methylations. When encountering potential methylation interference issues, a different isochimzomer can be attempted.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation Instructions for Use ： 1. Set up the single enzyme digestion reaction on ice as follows:ReagentPlasmid DNAUltrapure Waterto 20µl10X SgeⅠ Buffer2µlSubstrate DNAxµl (up to 0.5- 2µg)SgeI0.2-1µlTotal volume20µlNote: The reaction volume can be scaled up or down proportionally.a. Mix Gently by pipetting the reaction mixture up and down, or by flicking the reaction tube. Centrifuge briefly to collect the liquid at the bottom of the tube.b. Incubate at 37℃ for 1 hour preferably in a water bath, as the temperature is usually more stable. It is not recommended to exceed 1 hour for the incubation.c. (Optional) Terminate the reaction by heating at 80℃ for 20 minutes to inactivate the enzyme.2. When performing double or multiple enzyme digestion, the following principles can be referred for reaction setup based on the single enzyme digestion reaction provided above.a. Add 1μl of each restriction enzyme in the reaction. The reaction volume can be scaled up proportionally if needed.b. The total volume of restriction enzymes added in the reaction should not exceed 1/10 of the total reaction volume.c. If the optimal working temperature of different restriction enzymes used is different, start digestion with the enzyme that has a lower optimal temperature, then add other enzymes to perform the digestion at higher temperatures.