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来自 : 发布时间：2025-01-15

HomeProtocolsNEB PCR 克隆试剂盒的转化方案NEB PCR 克隆试剂盒的转化方案以下方案专为 NEB 10-beta 感受态大肠杆菌（NEB #C3019）设计，仅包含在 NEB PCR 克隆试剂盒（NEB #E1202）中。感受态细胞也可单独与 NEB #E1203 一起使用。重要 - 在使用不同的细胞株之前，请阅读有关感受态细胞要求的常见问题解答。将 50 μl 感受态细胞管在冰上解冻 10 分钟。添加 2 μl 连接反应;轻弹试管 4-5 次轻轻混合。在冰上孵育 20 分钟。在 42°C 下热休克 30 秒。在冰上冷却 5 分钟。加入 950 μl NEB™ 10-beta/Stable Outgrowth 培养基并放置在 37°C 下以 250 rpm 旋转或摇动 60 分钟（生长步骤）。注意：有关使用 NEB 10-beta 感受态大肠杆菌进行转化的进一步一般性讨论，请参阅 www.neb.com/ 上的转化方案C3019.克隆NEB PCR 克隆试剂盒 (NEB #E1202) 提供的高效细胞与高效菌株的区别仅在于它们的转化效率。此资源的链接产品类别：DNA 组装、克隆和诱变试剂盒产品应用：克隆和合成生物学相关产品：NEB® PCR 克隆试剂盒（无感受态细胞），NEB® PCR 克隆试剂盒HomeProtocolsNEB PCR 克隆试剂盒的电镀方案NEB PCR 克隆试剂盒的电镀方案通过轻弹或倒置彻底混合细胞，并将 50 μl 的 1 ml 生长物和 50 μl 的 1:10 稀释液扩散到 37°C 预热的琼脂平板上，其中含有 100 μg/毫升氨苄西林。对于 15 分钟的连接，传播两个体积的产物尤为重要，因为克隆插入物的选择最强，铺板密度 5\' exo-)工具与资源宣传册分子克隆技术指南选择工具A-tailing Selection Chart疑难解答指南末端修饰疑难解答指南法律信息

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