Tick the specs you need to submit them together; or enquire one by one. Cat. numbers are carried into the RFQ.
Q:TOPO 克隆和一步法克隆应用方面的区别?
A:TOPO 克隆主要用于构建克隆载体,应用于目的基因的复制,保存或测序等;一步法克隆可用于构建克隆或表达载体,点突变,多片段克隆等。
Q:TOPO 载体含有启动子吗?TOPO 载体是什么抗性?
A:没有启动子,因为TOPO 载体是克隆载体。氨苄抗性。
Q:载体能否提供全序列?客户实验能否用客户自己的载体?
A:a)可以提供全序列。
b)不能用客户的载体,试剂盒中已带有偶联TOPO 异构酶的载体。
Q:PCR 产物是否需要特殊处理?
A:无需特殊处理,但为了保证阳性率,最好进行胶回收纯化。
Q:平末端和 TA 克隆试剂盒的 Enhancer 能混合使用吗?
A:不建议混合使用。
Q:反应过夜可以吗?
A:不可以,TOPO 异构酶具有限制性内切酶活性,时间过长会重新切割重组质粒,建议反应时间不超过 5min。
Q:TOPO 载体最大可插入多大片段?能否插入多个片段?
A:a)验证过 5kb,一般片段越大,难度系数也会更高些。b)不能插入多片段,只能单个片段的插入。
Published citations
[1] Xu S, Jiang C, Lin R, et al. Epigenetic activation of the elongator complex sensitizes gallbladder cancer to gemcitabine therapy. J Exp Clin Cancer Res. 2021;40(1):373. Published 2021 Nov 25. doi:10.1186/s13046-021-02186-0(IF:11.161)
[2] Zhang W, Sun X, Wei X, Gao Y, Song J, Bai Y. Geography-Driven Evolution of Potato Virus A Revealed by Genetic Diversity Analysis of the Complete Genome. Front Microbiol. 2021;12:738646. Published 2021 Oct 1. doi:10.3389/fmicb.2021.738646(IF:5.640)
[3] Papolu PK, Ramakrishnan M, Wei Q, et al. Long terminal repeats (LTR) and transcription factors regulate PHRE1 and PHRE2 activity in Moso bamboo under heat stress. BMC Plant Biol. 2021;21(1):585. Published 2021 Dec 9. doi:10.1186/s12870-021-03339-1(IF:4.215)
[4] Fang Y, Tambo E, Xue JB, Zhang Y, Zhou XN, Khater EIM. Molecular Analysis of Targeted Insecticide Resistance Gene Mutations in Field-Caught Mosquitos of Medical Importance From Saudi Arabia. J Med Entomol. 2021;58(4):1839-1848. doi:10.1093/jme/tjab048(IF:2.278)
翌圣高保真酶,分子克隆的不二之选
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