摘要:Rnai最近由于RNA 干扰（RNA interference，RNA i）的发现使反义领域的研究增多。这种自然发生的现象最早是在秀丽线虫中发现的（1），是序列特异性地使转录后的基因沉默的有力机制。由于最近两年在RNA i领域取得的进步，已经有[阅读全文:]
摘要:The protocols listed here are for Drosophila cells in 6 well plates and our pre-aliquoted 384 well plates. RNAi experiments may be done in other size plates, just scale up or down.Adopted from Clemens[阅读全文:]
摘要:RNA干扰是最近发现的一种功能工具。当RNA导入一个细胞时，最终会引起细胞内互补mRNA的降解，从而导致基因功能活性的阻断。PTGS转录后基因沉默（posttranscriptionalgenesilencing）；一种首先在植物中确定，然后发现在动物中[阅读全文:]
摘要:为了分析特异siRNA 序列在基因活性上的效应，必须知道导入siRNA 的序列。这将要求设计合成寡核苷酸或者载体上的短的发夹RNA （siRNA ）序列，转染后检测基因的阻断。而dicer酶切的siRNA （d-siRNA ）导入细胞后，由于d-siRNA [阅读全文:]
摘要:Overview We design shRNA molecules with an algorithm. Our algorithm uses several criteria to rank potential 21mer targets within each human and mouse Refseq transcript. The algorithm applies a set of [阅读全文:]
摘要:基因敲除和转基因动物是功能缺失表型研究的主要方法。然而，获得这些动物是一个漫长而且费用昂贵的过程，同时很多基因的缺失会导致胚胎致死表型，使得基因敲除变得不再可能。灭活蛋白功能的方法包括结构域阴性突变构建和[阅读全文:]
摘要:Long double-stranded RNAs (dsRNAs; typically >200 nt) can be used to silence the expression of target genes in a variety of organisms and cell types (e.g., worms, fruit flies, and plants). Upon introd[阅读全文:]
摘要:1. Add the following to a sterile microfuge tube on ice:-1 ug linearized plasmid DNA.-2 ul Dig RNA Labeling Mix, 10X concentrate-2 ul 10X transcription buffer-1-2 ul RNase inhibitor (DNase free)-add D[阅读全文:]
摘要:Northern BlotPreparation of Formaldehyde Agarose Gel The gel conditions (1% agarose, 1X MOPS, 6.3% formaldehyde) are designed for ~4 hours of electrophoresis. If longer times are necessary, the formal[阅读全文:]
摘要: OverviewWith this protocol, transcripts that were initiated from specific genes by RNA polymerases prior to permeabilization can be measured. Instead of a nuclear extract, permeabilized cells are use[阅读全文:]