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產(chǎn)品信息

貨號	名稱	規(guī)格	單價/元
TSM99K2	EZ-Tn5? <Kan-2>Tnp Transposome? Kit/EZ-Tn5? <Kan-2>Tnp轉(zhuǎn)座試劑盒	10 rxns	6775
TY0261H	TypeOne? Restriction Inhibitor/TypeOne?限制抑制劑	100 μg	2469
TSM08KR	EZ-Tn5? <R6Kγori/Kan-2>Tnp Transposome? Kit	10 rxns	7232

產(chǎn)品簡介

      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轉(zhuǎn)座酶和EZ-Tn5 <R6Kγori/ KAN-2>轉(zhuǎn)座子之間形成的穩(wěn)定復合物。EZ-Tn5 <R6Kγori/ KAN-2>轉(zhuǎn)座子包含R6Kγ條件復制起點（R6Kγori）和在大腸桿菌中有功能的Tn903卡那霉素抗性基因（KanR），側(cè)翼是19堿基對活躍的Mosaic End（ME）EZ -Tn5轉(zhuǎn)座酶識別序列。EZ-Tn5轉(zhuǎn)座子可以電穿孔進入活細胞，在宿主細胞的環(huán)境中，Mg2 +激活EZ-Tn5轉(zhuǎn)座酶，從而將EZ-Tn5轉(zhuǎn)座子隨機插入宿主的基因組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使該轉(zhuǎn)座子可用于“營救”隨機插入轉(zhuǎn)座子的基因組DNA區(qū)域。 第2頁概述了營救克隆過程。首先將轉(zhuǎn)染EZ-Tn5 <R6Kγori/ KAN2>的轉(zhuǎn)座子的基因組DNA純化，然后進行片段化，自連接，最后轉(zhuǎn)化為表達pir的大腸桿菌宿主。 基因產(chǎn)物（“ pi”蛋白）。在含卡那霉素的平板上選擇時，只有包含<R6Kγori/ KAN-2>轉(zhuǎn)座子的細胞才能生長。

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.
    試劑盒中提供了未標記的正向和反向轉(zhuǎn)座子特異性引物。 這些引物可用于雙向DNA測序或靶基因組DNA或救援克隆中轉(zhuǎn)座子插入位點的作圖。

組成成分：
? EZ-Tn5 <R6Kγori/KAN-2> Tnp Transposome (0.1pmol/μL)：儲存在-20°C
? KAN-2 FP-1 Forward Primer (50   μM)：儲存在-20°C
? R6KAN-2 RP-1 Reverse Primer (50 μM)：儲存在-20°C
? Nuclease-Free Water, Sterile：儲存在-20°C
數(shù)據(jù)：

Figure 1. An EZ-Tn5 Transposome is the stable complex formed by   EZ-Tn5 Transposon with EZTn5 Transposase in the absence of MG2+.
圖1. EZ-Tn5轉(zhuǎn)座體是在沒有MG2 +的情況下由EZ-Tn5轉(zhuǎn)座子與EZTn5轉(zhuǎn)座酶形成的穩(wěn)定復合物。

Figure 2. An EZ-Tn5 Transposome Complex can be electroplated into living cells where it randomly insert the transposon insertion site can be host’s genomic DNA. The EZ-Tn5 Transposon insertion site can be analyzed by a variety of methods.
圖2.可以將EZ-Tn5轉(zhuǎn)座體復合物電轉(zhuǎn)到活細胞中，在其中隨機插入轉(zhuǎn)座子插入位點，可以是宿主的基因組DNA。 EZ-Tn5轉(zhuǎn)座子插入位點可以通過多種方法進行分析。
相關(guān)產(chǎn)品：
? TypeOne Restriction Inhibitor（# TY0261H）
? TransforMax EC100D pri+ Electrocompetent  E.coli（# ECP09500 ）
? TransforMax EC100D pir-116 Electrocompetent  E.coli（# EC6P095H）

參考文獻：
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.