Hieff Mut® Multi Site-Directed Mutagenesis Kit 多点突变试剂盒
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:多点突变试剂盒与定点突变试剂盒有什么区别？
A:多点突变试剂盒可一步实现目标质粒上 3-5 个不连续位点的定点突变，而定点突变试剂盒是
适合 1 或 2 个不连续位点的定点突变。
如：3 个连续的碱基突变算 1 个位点的突变，故定点突
变试剂盒即可（相距超过 50 bp 为 1 个不连续位点）。
Q:点突变为什么要选择甲基化的载体？
A:点突变过程需要去除模板质粒，防止假阳性。利用的原理就是限制性内切酶DpnI 能消化甲基化的模板，若是非甲基化，则无法去除模板。
Q:对于 PCR 模板质粒什么要求？
A:a）PCR 模板应为甲基化质粒，应使用甲基化酶无缺陷型的宿主菌扩增原始质粒（如DH5α， TOP10，JM109 等）。
b）应尽量使用新鲜制备的质粒作为模板，因长期放置，反复冻融可能会导致模板质粒断裂， 开环或降解。
Q:质粒反向扩增时引物如何设计？
A:正反向扩增引物 5’端包含至少 20 bp 反向互补区域，各引物非互补区域长度至少为 20 bp。互补区尽量选择无重复序列，GC 含量尽量在 40%-60%范围内。
已发表文献
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