计算溶液所需的质量、体积或浓度。
BioReagent,EnzymoPure™,for DNA and RNA applications,PCR试剂,分子生物学级,用于DNA合成,用于体外转录 级 ,适用于对基线干扰要求严格的色谱和分析工作流程。
-20°C储存,避免反复冻融。超低温运输 。请查阅批次 COA 获取详细规格。
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在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
储存缓冲液: 10mM Tris-HCl, 50mM KCl, 1mM DTT, 0.1mM EDTA, 200µg/ml Recombinant Albumin, 50% Glycerol, pH 7.4
NruI可识别 TCG/CGA 回文序列,酶切后形成平末端,其同裂酶有 Bsp68I、BtuMI、RruI。NruI 和 10×Cut Reaction Buffer 均添加了重组白蛋白(rAlbumin),确保了产品的安全性及稳定性。
产品组分
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来源
E.coli
酶切位点
5’…TCG↓CGA…3’
3’…AGC↑GCT…5’
酶活定义
在 50 μl 总反应体系中,37℃条件下,1 h内酶切 1 μg λDNA 所需的酶量定义为 1 个活力单 位(U)。
产品使用步骤
(1)按照下表所示配置混合溶液
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(2) 轻柔吸打混匀,切勿振荡涡旋,然后瞬时离心。
(3) 37°C孵育 30 min。
注:*Nrul 最后加入体系中,一般 50 μl 反应体系加入1μl 的Nrul,也可根据实验进行调整,建议酶的加入量不超过总体系的10%,避免产生星活性。
注意事项
(1) Nrul 的甲基化敏感性:
Dam 甲基化:甲基化与酶切位点可能重叠导致切割受损
Dcm 甲基化:不敏感
CpG 甲基化: 阻断切割
(2) 在梅切反应前,尽量确保DNA不含苯酚、氯仿、酒精、EDTA 等杂质,以免影响酶切效果。
(3) 本产品仅作科学研究使用,不得用于其它用途。
Storage buffer: 10mM Tris-HCl, 50mM KCl, 1mM DTT, 0.1mM EDTA, 200µg/ml Recombinant Albumin, 50% Glycerol, pH 7.4
NruI can recognize the TCG/CGA palindromic sequence, and form a flat terminal after enzyme digestion. The isozymes of NRUI include Bsp68I, BtuMI and RruI. Recombinant albumin was added to both NruI and 10×Cut Reaction Buffer to ensure the safety and stability of the product.
Product composition
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Source
E.coli
Restriction Enzyme CuttingSite
5’…TCG↓CGA…3’
3’…AGC↑GCT…5’
Enzyme Activity Definition
In a total reaction system of 50 μl, the amount of enzyme required for enzyme digestion of 1 μg λDNA within 1 h at 37℃ was defined as one active single site (U).
Protocol
(1) Configure the reaction system, as shown in the following table
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(2) Gently mix, do not oscillate the vortex, and then instantaneous centrifugation.
(3) Incubation at 37°C for 30 min.
Note: *Nrul is finally added to the system, generally 50 μl reaction system to add 1μl of Nrul, can also be adjusted according to the experiment, it is recommended that the amount of enzyme added does not exceed 10% of the total system to avoid star activity.
Cautions
(1) Methylation sensitivity of Nrul:
Dam methylation: Methylation and cleavage sites may overlap resulting in impaired cleavage
Dcm methylation: insensitive
CpG methylation: Blocking cleavage
(2) Before the Meche reaction, try to ensure that the DNA does not contain impurities such as phenol, chloroform, alcohol, EDTA, so as not to affect the effect of enzyme digestion.
(3) This product is for scientific research only and shall not be used for other purposes.
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