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摘要:Selection of siRNA duplexes from the target mRNA sequenceUsing Drosophila melanogaster lysates (Tuschl et al. 1999), we have systematically analyzed the silencing efficiency of siRNA duplexes as a fun[阅读全文]
摘要:The siRNA user guide(revised May 6, 2004)Selection of siRNA duplexes from the target mRNA sequenceUsing Drosophila melanogaster lysates (Tuschl et al. 1999), we have systematically analyzed the silenc[阅读全文]
摘要:Our current strategy with siRNA is to synthesis relatively small amounts enzymatically and use these to test for efficiency by western blotting. This allows us to test several candidate siRNAs quite c[阅读全文]
摘要:RNAi target selection rules : Targeted regions on the cDNA sequence of a targeted gene should be located 50-100 nt downstream of the start codon (ATG). Search for sequence motif AA(N19 )TT or NA(N21 )[阅读全文]
摘要:越来越多的研究人员开始采用小分子干扰RNA(small interfering RNAs,siRNAs)来抑制特定的哺乳动物基因表达。siRNA是一种短片断双链RNA分子,能够以同源互补序列的mRNA为靶目标降解特定的mRNA,这个过程就是RNA干扰途径(R[阅读全文]
摘要:Using siRNA for gene silencing is a rapidly evolving tool in molecular biology. There are several methods for preparing siRNA, such as chemical synthesis, in vitro transcription, siRNA expression vect[阅读全文]
摘要:siRNA Design RNAi target selection rules :Targeted regions on the cDNA sequence of a targeted gene should be located 50-100 nt downstream of the start codon (ATG). Search for sequence motif AA(N19 )TT[阅读全文]
摘要: A.普通阴性对照1.siRNA实验应该有阴性对照;2.通用阴性对照为与目的基因的序列无同源性的普通阴性对照;3.Scrambled阴性对照和选中的siRNA序列有相同的组成,但是和mRNA没有明显的同源性;4.阴性对照需要确定和目的靶细[阅读全文]
摘要:将制备好的siRNA,siRNA表达载体或表达框架转导至真核细胞 中的方法主要有以下几种: 1.磷酸钙共沉淀将氯化钙,RNA(或DNA )和磷酸缓冲液混合,沉淀形成包含DNA 且极小的不溶的磷酸钙颗粒。磷酸钙-DNA 复合物粘附到细胞 膜[阅读全文]
摘要:1.希望软件可以在 web 上使用。 即用户可以访问我们的网站, 输入基因代码, 即可自行在网上设计 siRNA Oligo 。2.可参照的软件形式: 见如下网站: www.dharmacon.comwww.ambion.comwww.qiagen.com3.希望能就每一个所指定[阅读全文]
摘要:siRNA Design RNAi target selection rules :Targeted regions on the cDNA sequence of a targeted gene should be located 50-100 nt downstream of the start codon (ATG). Search for sequence motif AA(N19 )TT[阅读全文]
摘要:Ambion and Applied Biosystems have joined forces to provide a complete convenient, solution for performing gene silencing experiments and validating the results by real-time RT-PCR. Ambion's Silencer&[阅读全文]
摘要:在活体RNAi关键点:类似药物属性的引入,如稳定性、细胞内的传送、组织的生物可溶性进入合成的siRNA。授予siRNA类似药物的属性化学稳定性的胆固醇共聚的siRNA 显著的改善了其在体内和体外的药物属性。化学稳定性增强的方法[阅读全文]
摘要:1. 对每个基因设计并检测两到四个siRNA 序列为了找到潜在靶位点,扫描靶基因中的AA序列。记录每个AA 3’端19个核苷酸作为潜在siRNA 靶位点。潜在靶位点需通过GENBANK数据库的BLAST分析,去除那些与其它基因明显同源[阅读全文]
摘要:siRNA Database Searchable database of Silencer ™ Validated and Pre-designed siRNAs to >34,000 human, mouse, and rat targets. All siRNAs in the database have been designed for maximum potency and[阅读全文]
摘要:siRNA序列设计注意事项:选择以AA开头的19-23nt长的序列;靶定序列从起始密码子下游100个nt处开始,到终止密码子上游100nt处结束。本篇文章来源于网络,如有异议请联系我们,我们将在3个工作日内作出处理。[阅读全文]
摘要:化学合成的siRNA 与生物合成siRNA 相比,有以下优点:?操作简便,研究人员得到合成好的siRNA 后,进行简单的稀释处理即可实验。?转染效率高,相对于分子量较大的DNA 质粒,其转染效率高。?对细胞的或者组织的毒副作用小。?可[阅读全文]
摘要:一、体外制备1、化学合成2、体外转录 3、用RNase III 消化长片断双链RNA制备siRNA二、体内表达1、siRNA表达载体或者病毒载体在细胞中表达siRNAs2、PCR制备的siRNA表达框在细胞中表达RNAi可以直接用于疾病相关基因的抑制,从[阅读全文]