OK Clon DNA 连接试剂盒 II

产品货号：AG11807 / AG11808

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规格

10 rxns / 10 μl

50 rxns / 10 μl

清除

价格范围：¥580 至 ¥2,400

产品描述

OK Clon DNA Ligation Kit II 是一种准确、快速、高效的定向克隆试剂盒。本试剂盒可以在 10 分钟内，快速高效地将一段或多段 DNA 片段定向、无缝的克隆到任意载体的任意位置，而不被酶切位点限制。本产品是在 OK Clon DNA Ligation Kit（Code. AG11802 / AG11803）的基础上进行了优化，适用范围更广，尤其适合大载体、大片段及多片段的连接。DNA 连接反应是基于 OK Clon Enzyme 3’- 5’ 端的外切酶活性，从线性化 DNA 链的 3’ 末端切割核苷酸形成粘性末端，由此，片段与载体末端能够形成 15 ~ 20 bp 的互补序列，通过退火可以准确、高效的将片段与载体连接。只需在设计引物时将与载体 5’ 端互补的序列添加在引物 5’ 端，通过该引物扩增目的片段，无需对 PCR 片段进行限制酶切、磷酸化等处理，就可将目的片段插入载体中。

产品优势

1. 可将片段插入载体的任意位置，而不受酶切位点限制；
2. 反应迅速，50℃ 10 min 即可完成连接反应；
3. 适用范围更广，适用于各种大小的载体与片段的连接；特别适合大载体、大片段及多片段的连接；
4. 插入片段不需酶切处理，PCR 扩增完成后只需要进行纯化处理便可直接进行连接反应。

产品组成

组分	AG11807	AG11808
2.5 X OK Clon Master Mix	40 μl	100 μl x 2 pc

保存与运输

保存温度：-20℃保存
运输温度：干冰运输或-20℃冰袋运输

示例一

使用本产品搭配 Linearized Control Vector & Insert （for OK Clon）（Code No.AG11814）中的线性化载体及片段进行克隆实验：
① 按照下表配制反应体系：

组分名称	实验组	阴性对照	*阳性对照^1**
2.5 X OK Clon Master Mix	4 μl	4 μl	4 μl
Linearized Control vector（50 ng / μl）	1 μl	1 μl	/
2 kb Positive Control Insert（80 ng / μl）	1 μl	/	/
RNase free water	4 μl	5 μl	Up to 10 μl

*1：阳性对照仅加入 100 pg 环状的 pUC19 质粒。不加线性化载体与片段。

② 将所有组别转化至 E.coli DH5α Competent cells（Code No. AG11806），培养的菌悬浮液稀释 3 倍后，100μl 涂布平板（本实验使用 LB 培养基，加入了 Amp 抗生素、IPTD，以及 X-Gal，用于蓝白斑筛选）。结果如下图所示，实验组的菌落均匀，得到数百个单克隆菌落。