产品信息

产品简介

      The EZ-Tn5™ <R6Kγori/KAN-2>Tnp Transposome™ is the stable complex formed between the EZ-Tn5 Transposase enzyme and the EZ-Tn5 <R6Kγori/KAN-2> Transposon.The EZ-Tn5 <R6Kγori/KAN-2> Transposon contains an R6Kγconditional origin of replication (R6Kγori) and the Tn903 kanamycin resistance gene (KanR) that is functional in E. coli, flanked by hyperactive 19-basepair Mosaic End (ME) EZ-Tn5 Transposase recognition sequences. The EZ-Tn5 Transposome can be electroporated into living cells where the EZ-Tn5 Transposase is activated by Mg2+ in the host’s cellular environment resulting in random insertion of the EZ-Tn5 Transposon into the genomic DNA of the host.
      EZ-Tn5™<R6Kγori/ KAN-2> Tnp Transposome™是EZ-Tn5转座酶和EZ-Tn5 <R6Kγori/ KAN-2>转座子之间形成的稳定复合物。 EZ-Tn5 <R6Kγori/ KAN-2>转座子包含R6Kγ条件复制起点（R6Kγori）和在大肠杆菌中有功能的Tn903卡那霉素抗性基因（KanR），两侧为19碱基对活跃的Mosaic End（ME）EZ -Tn5转座酶识别序列。 EZ-Tn5转座子可以电穿孔进入活细胞，在宿主细胞的环境中，Mg2 +激活EZ-Tn5转座酶，从而将EZ-Tn5转座子随机插入宿主的基因组DNA中。
The R6Kγori makes this transposon useful for “rescue cloning”of the region of genomic DNA into which the transposon has been randomly inserted. An overview of the rescue cloning process is presented on page 2. Genomic DNA transposed with the EZ-Tn5 <R6Kγori/KAN2> Transposon is   first purified and then fragmented, self-ligated and finally transformed into   an E. coli host that expresses the pir gene product (the “pi” protein).When selected on kanamycin-containing plates, only the cells containing the <R6Kγori/KAN-2> Transposon will grow.
      R6Kγori使该转座子可用于“营救”随机插入转座子的基因组DNA区域。 第2页概述了急救克隆过程。首先将转染EZ-Tn5 <R6Kγori/ KAN2>的转座子的基因组DNA纯化，然后进行片段化，自连接，最后转化为表达pir的大肠杆菌宿主。 基因产物（“ pi”蛋白）。在含卡那霉素的平板上选择时，只有包含<R6Kγori/ KAN-2>转座子的细胞才能生长。
Unlabeled forward and reverse transposon-specific primers are supplied in the kit. These primers can be used for bidirectional DNA sequencing or mapping of transposon insertion sites in target genomic DNAs or rescue clones.
     试剂盒中提供了未标记的正向和反向转座子特异性引物。 这些引物可用于双向DNA测序或靶基因组DNA或救援克隆中转座子插入位点的作图。
组成成分：
? EZ-Tn5 <R6Kγori/KAN-2> Tnp Transposome (0.1   pmol/μL)：储存在-20°C
? KAN-2 FP-1 Forward Primer (50   μM)：储存在-20°C
? R6KAN-2 RP-1 Reverse Primer (50 μM)：储存在-20°C
? Nuclease-Free Water, Sterile：储存在-20°C
数据：

参考文献：
1.Hoffman, L.M. and Jendrisak, J. (1999) Epicentre Forum, 6 (3), 1.
2.Goryshin, I.Y. et al., (2000) Nat. Biotechnol. 18, 97.
3.Metcalf, W.W. et al.,   (1994) Gene, 138, 1.