Hieff Clone® Zero TOPO-TA Cloning Kit 零背景TOPO-TA克隆试剂盒
勾选需要的规格可一次提交；也可以逐个询价。货号会一并带入询价单。
Q：从质粒上酶切的产物可直接连接 Topo 载体吗？为什么？
A：Topo 载体进行连接反应的时候需要 5`羟基端和 Topo 酶发生反应。PCR 产物 5`羟基端没有磷酸化， 可以直接进行 TOPO 克隆。但是，酶切产物 5`端为磷酸化状态，可以直接进行 TA 克隆的，但是不能进行 Topo 克隆。如果需要，可以用牛小肠碱性磷酸酶（CIP）进行去磷酸化处理后连接 Topo 载体。
Q：连接反应体系和反应时间以及温度对连接效率的影响大吗？
A：建议反应体系应控制在 10μl 内，体系太小连接会受到影响。如果目的基因浓度太低，建议浓缩之后再连接，以提高连接成功率。插入片段使用量参考说明书。一般 1-2min 即可完成连接反应，若想得到更多克隆，可适当延长至 5min，但不能超过 5min。室温反应即可。
Q：如何选择合适的载体？
A：若 PCR 产物为 Taq 酶扩增则选择 TOPO-TA 克隆载体系列，若 PCR 产物为高保真酶扩增则选择 Blunt 载体系列，Simple 载体不含多克隆位点区，可根据需求在 PCR 扩增时添加自己需要的酶切位点。
Q：若是储存了很久的 PCR 产物还能用来做克隆吗？
A：建议采用新鲜的 PCR 产物会使连接效率更高一些，若是-20℃保存的一周内还可以使用。请避免PCR 产物的反复冻融和降解。
Q：PCR 扩增产物需要纯化吗？
A：先对 PCR 产物跑胶鉴定，若是 PCR 产物电泳条带单一、浓度较高且无引物二聚体可直接进行连接反应。若是有杂带或引物二聚体建议纯化之后再进行连接反应。
已发表文献
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