ReadiLink Cy5 缺口平移 dsDNA 标记试剂盒
| Ex (nm) | 651 | Em (nm) | 670 |
| 分子量 | N/A | 溶剂 | - |
| 存储条件 | - |
ReadiLink Cy5 缺口平移 dsDNA 标记试剂盒提供了一种简单有效的方法，可以使用明亮且光稳定的 Cy5染料标记双链 DNA 样本。标记试剂盒为 DNA 标记所需的完整工作流程提供了所有必要的试剂。该方法使用 DNAse 和 DNA 聚合酶的组合来切割 DNA 螺旋的一条链，Cy5染料与之结合。此外，该试剂盒允许用户通过调整 Cy5-dUTP 偶联物与 dTTP 的比例来优化掺入和产品大小。它与多种样品材料兼容，包括人工染色体 (BAC) DNA、人类基因组 DNA、纯化的 PCR 产物、超螺旋和线性化质粒 DNA。得到的 Cy5标记 DNA 可用于多种分子生物学技术，例如荧光原位杂交 (FISH)。百萤生物是AAT Bioquest的中国代理商，为您提供优质的ReadiLink Cy5缺口平移 dsDNA 标记试剂盒。
样品实验方案
简要概述
- 准备 DNA 样本
- 向试管中加入试剂
- 短暂混合并离心
- 在 15°C 下孵育 60 分钟
- 将反应置于冰上，然后加入停止溶液并在 65°C 下加热
- 使用前在冰上放置 5 分钟或在 4°C 下储存
- 纯化标记的 DNA
注：在开始实验之前，解冻所有成分。在开始标记过程之前，将所有试剂短暂涡旋至底部。
实验步骤
表 1. 各反应每管试剂组成
| 成分 | 规格 |
| DNA样本 | 1 µg DNA 在无核酸酶水中稀释至终体积 34 µL |
| Nick Translation 缓冲液 | 5 µL |
| dNTP 混合物 | 5 µL |
| dTTP | 2 µL |
| Cy5-dUTP 工作溶液 | 2 µL |
| DNA聚合酶I | 1 µL |
| DNA酶I | 1 µL |
| 总容积 | 50 µL |
可以优化孵育时间以获得更好的标记。 更长的孵育时间将有助于更多的标记，但可能会缩短终产品的尺寸。
1.向干净的（无核酸酶）0.5 mL 微型离心管或 0.2 mL PCR 管中，按表 1 中所示的顺序添加试剂。
2.通过短暂的涡旋和短暂的离心小心混合试剂。
3.将反应在 15°C 下孵育 60 分钟。
4.孵育后，将反应置于冰上。
5.要终止反应，请添加 5 µL 停止溶液并将样品加热至 65 °C。
6.使用前在冰上放置 5 分钟或在 4°C 下储存。
7.纯化标记的 DNA。
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