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慢病毒包装试剂盒
发布日期:2015-10-03 19:39:05 浏览次数 :
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Lenti-TSB™ 慢病毒包装试剂盒Transheep的的Lenti-TSB™ 表达包装系统包括优化的慢病毒包装质粒
  Lenti-TSB™ 慢病毒包装试剂盒    
           
  Transheep的的Lenti-TSB™ 表达包装系统包括优化的慢病毒包装质粒,有多种Tag及启动子可供选择copGFP阳性对照质粒,Namipo-T新型转染试剂等。该系统与的Transheep的ORF、shRNA克隆配套使用保证更高滴度和更高表达水平,同时为您降低可观的实验成本。    
           
    内容 价格    
  慢病毒空载 丰富的载体可供选择,包括不同的荧光、抗性、启动子 800元起    
  模版合成 如果您没有目的基因的模版,我们可以帮您合成任意序列并克隆到您选定的载体 低至1元/bp    
  载体构建        构建任意您感兴趣的基因到您选择的载体,包括LncRNA等,最大容量为6kb 800元起    
  表达检测 构建好的载体少量抽提,转染工具细胞进行目的基因表达检测,保证您收到的载体可高效表达 500元起    
  包装体系  经过优化的高纯度包装质粒体系,按推荐比例与穿梭质?;旌霞纯?/td> 500元起    
   质粒抽提  去内毒素高纯度质粒是慢病毒包装的必备条件,Transheep可提供质粒制备及专用的抽提试剂盒 500元起    
  转染试剂 Namipo-T™ ,纳米材料293t专用新型转染试剂,效率可达90%以上,几乎无毒,更利于出毒 800元起     
  包装及滴度检测 提供包装好的纯化慢病毒,保证滴度达到要求 2000元起    
           
实验流程     
  构建目的基因的慢病毒载体,制备慢病毒穿梭质粒及其辅助包装原件载体质粒,分别进行高纯度无内毒素抽提,使用Namipo-T共转染 293T 细胞,培养 48和 72h 后,分别收集富含慢病毒颗粒的细胞上清液,病毒上清液通过超离心浓缩病毒。    
实验材料     
           
(1)慢病毒载体、包装细胞和菌株    
  Transheep的的Lenti-TSB™ 表达包装系统,可根据您的需求现在最适合的表达载体,多种启动子可选,CMV、PGK 启动子在常规肿瘤细胞有极高的表达能力,而 MSVC、EF1 启动子则可满足干细胞中过表达基因的需求。Puro、Neo、 抗性,copGFP、tRFP、Luc 等多种抗性基因与荧光标记的选择搭配。
可实现多种基因的共转化。出更高的出毒效率。
   
       
货号 类别 载体名称 荧光标记 抗性 蛋白标签
TSE01 慢病毒OE pTSBOE-CMV-MSC-3flag-EF1-copGFP 绿色 Flag
TSE02 慢病毒OE pTSBOE-CMV-MSC-3flag-EF1-copGFP-F2A-Puro 绿色 Puro Flag
TSE03 慢病毒OE pTSBOE-CMV-MSC-3flag-EF1-tRFP-F2A-Puro 红色 Puro Flag
TSE04 慢病毒OE pTSBOE-CMV-MSC-3flag-EF1-copGFP-F2A-Neo 绿色 Neo Flag
TSE05 慢病毒OE pTSBOE-CMV-MSC-3flag-EF1-tRFP-F2A-Neo 红色 Neo Flag
TSE06 慢病毒OE pTSBOE-CMV-MSC-EF1-Puro Puro
TSE07 慢病毒OE pTSBOE-CMV-MSC-IRES-Neo Neo
TSE08 慢病毒SH pTSBSH-U6-MSC-CMV-ZsGreem-PGK-Puro 绿色 Puro  
TSE09 慢病毒SH pTSBSH-U6-MSC-CMV-ZsGreem-PGK-Neo 绿色 Puro  
TSE10 慢病毒SH pTSBSH-U6-MSC-CMV-tRFP-PGK-Puro 红色 Neo  
TSE11 慢病毒SH pTSBSH-U6-MSC-CMV-tRFP-Neo 红色 Neo  
TSE12 慢病毒SH pTSBSH-U6-MSC-CMV-Neo Neo  
TSE13 慢病毒SH pTSBSH-U6-MSC-CMV-Puro Puro  
TSE14 慢病毒SH pTSBSH-H1-MSC-FE1-Puro-copGFP 绿色 Puro  
TGL004 慢病毒 pTsLVGL-MCS-Minp-Luc2-EF1-Puro LUC2 Puro 转录因子、启动子验证
TGL005 慢病毒 pTsLVGL-CMV-Luc2-EF1-Puro LUC2 Puro UTR验证,启动子验证
     
(2) 包装体系    
  Transheep的 packing mix是我们经过优化过的包装体系,可以按照穿梭质粒与包装体系(1+基因大小kb)/1的比例混合即可,Lenti-TSB™也可以使用通用的包装细胞进行包装。    
(3)菌株大肠杆菌菌株    
  推荐使用top10作为质粒扩增及构建的菌株。如果基因复杂可考虑使用stbl3菌株,保真度要高一些,正常情况下使用top10引起载体突变的概率不大。    
(4)包装细胞      
  TSB-293Tmax™ 细胞系转染效率高,能够实现病毒蛋白高水平的表达,表现出更高的出毒效率。    
(5)试剂耗材        
  慢病毒包装所用的质粒必须去内毒素处理,培养耗材及血清也是影响慢病毒出毒的一大因素,市场上的试剂假冒产品较多,可咨询客户购买适合包装所用的相关产品。    
           
实验流程    
  请参考TSB慢病毒包装试剂盒使用说明    
       
参考文献    
  The antihelminthic drug niclosamide effectively inhibits the malignant phenotypes of uveal melanoma in vitro and in vivo;Theranostics. 2017; 7(6): 1447–1462.
Published online 2017 Apr 3. doi:  10.7150/thno.17451
   
  USP19 suppresses cellular type I interferon signaling by targeting TRAF3 for deubiquitination;Published Online:10 Apr 2017https://doi.org/10.2217/fmb-2017-0006    
  Upregulation of the Non-Coding RNA OTUB1-isoform 2 Contributes to Gastric Cancer Cell Proliferation and Invasion and Predicts Poor Gastric Cancer Prognosis;Int J Biol Sci. 2016; 12(5): 545–557.
Published online 2016 Mar 3. doi:  10.7150/ijbs.13540
   
  Pristimerin effectively inhibits the malignant phenotypes of uveal melanoma cells by targeting NF?κB pathway;Published online on: July 25, 2017   https://doi.org/10.3892/ijo.2017.4079    
  Multifunctional nanoparticles for co-delivery of paclitaxel and carboplatin against ovarian cancer by inactivating the JMJD3-HER2 axis;The article was received on 21 Jun 2017, accepted on 31 Jul 2017 and first published on 03 Aug 2017    
  Long non-coding RNA LOC389641 promotes progression of pancreatic ductal adenocarcinoma and increases cell invasion by regulating E-cadherin in a TNFRSF10A-related manner;https://doi.org/10.1016/j.canlet.2015.12.010    
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