摘要:The RNA in situ procedure described below is based on the protocol developed by Tautz and Pfeifle (Chromosoma 98 (1989), p81), but adapted to allow the screening of 96 RNA probes on whole-mount Drosop[阅读全文:]
摘要:N.B: Gloves should be worn at all times during preparation of in vitro RNA. All solutions should be RNase-free i.e. made with DEPC water if home-made or bought specifically to use for RNA work. You wi[阅读全文:]
摘要:First strand of cDNA synthsis1.Add 2 μl of Not I primer-adapter to a sterile 1.5 ml RNAase free tube, add 4.5 μl mRNA of molt4 cells (~5.2 ug), add 0.5 μl DEPC-H2O.2. Heat the mixture at 70℃ [阅读全文:]
摘要:General Guidelines siRNA targeted sequence is usually 21 nt in length. Avoid regions within 50-100 bp of the start codon and the termination codon Avoid intron regions Avoid stretches of 4 or more bas[阅读全文:]
摘要:A novel approach for evaluating the efficiency of siRNAs on protein levels in cultured cells. Wu W, Hodges E, Redelius J, Hoog C. Nucleic Acids Res. 2004 Jan 22;32(2):E17. Center for Genomics and Bioi[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: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: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:The use of small interfering RNAs (siRNAs) to induce targeted gene silencing in mammalian cells offers researchers a powerful tool for analyzing gene function. Ideally one would like to work with indi[阅读全文:]