TheTRCshRNADesignMethodsandRules2
Score the Candidates Sequences for SpecificityWe are forced to balance the prediction of knockdown efficiency against the desire to minimize interaction with off-target genes, without a clear understanding of just how to predict off-target "hits". We calculate a "specificity score" to promote candidates without obvious off-target transcripts. Each candidate is compared by BLASTN to two distinct ab......阅读全文
The－TRC－shRNA－Design－Methods－and－Rules
OverviewWe design shRNA molecules with an algorithm. Our algorithm uses several criteria to rank potential 21mer targets within each human and mouse R
The－TRC－shRNA－Design－Methods－and－Rules2
Score the Candidates Sequences for SpecificityWe are forced to balance the prediction of knockdown efficiency against the desire to minimize interacti
The－TRC－shRNA设计方法与原则
OverviewWe design shRNA molecules with an algorithm. Our algorithm uses several criteria to rank potential 21mer targets within each human and mouse R
Rules－of－siRNA－design－for－RNA－interference－(RNAi)
General GuidelinessiRNA targeted sequence is usually 21 nt in length.Avoid regions within 50-100 bp of the start codon and the termination codonAvoid
BLOCKiT－RNAi－Designer
Allows you to design synthetic siRNA, Stealth RNA, or shRNA molecules from nucleotide target sequences, or convert an siRNA molecule sequence into a S
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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).S
设计shRNA教程
一、认识shRNA： 在研究基因功能中，RNAi由于可以特异性地使基因沉默或表达量降低而成为生物实验中的有力工具。其中，shRNA是可以克隆至表达载体并表达siRNA双链的DNA分子。今天给大家分享两种能快速设计shRNA的方法。 二、shRNA的设计: 1. Sigma网站Sigma公司做了一个针
Northern－protocol(RULES－FOR－RNA－WORK)
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shRNA干扰载体构建
产品技术背景pRI系列载体是基于III类RNA聚合酶启动子：人类H1启动子的专用于哺乳动物细胞RNA干扰的载体。H1启动子在哺乳动物细胞内合成类似siRNA分子的小分子RNA。由于H1启动子有精确的转录起始位点和终止信号，H1启动子转录产物精确生成人工设计的shRNA，shRNA经过RISC剪切后形
shRNA干扰载体构建
产品技术背景pRI系列载体是基于III类RNA聚合酶启动子：人类H1启动子的专用于哺乳动物细胞RNA干扰的载体。H1启动子在哺乳动物细胞内合成类似siRNA分子的小分子RNA。由于H1启动子有精确的转录起始位点和终止信号，H1启动子转录产物精确生成人工设计的shRNA，shRNA经过RISC剪切后形
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1. 载体取2-5ug，在25ul体系中，用2.5ul的酶切30min-1h，回收载体：载体:---2ul（2-5ug）酶---------2.5ulBuf--------2.5ul水---------18ul回收的时候希望浓度高点。2. Oligo退火形成双链若为2OD=5.4nmol的话，则每个
siRNA－Design－Guidelines
Using siRNA for gene silencing is a rapidly evolving tool in molecular biology. There are several methods for preparing siRNA, such as chemical synthe
手把手教你设计shRNA
一、认识shRNA： 在研究基因功能中，RNAi由于可以特异性地使基因沉默或表达量降低而成为生物实验中的有力工具。其中，shRNA是可以克隆至表达载体并表达siRNA双链的DNA分子。今天给大家分享两种能快速设计shRNA的方法。 二、shRNA的设计: 1. Si
体内表达shRNA的设计
1.克隆到shRNA 表达载体中的shRNA 包括两个短反向重复序列，中间由一茎环（loop）序列分隔的，组成发夹结构，由polⅢ启动子控制。随后在连上5-6个T作为RNA 聚合酶Ⅲ的转录终止子。2.两个互补的寡核苷酸两端须带有限制性酶切位点。3.Stratagene发现29个寡核苷酸较之原先推荐的
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Adapted from TaqMan® One-Step RT-PCR Master Mix Reagents Kit InstructionDesign of Probes Keep the G-C content in the 20 to 80% range.Avoid runs of an
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References Albert, J., and Fenyo, E.M. 1990. Simple, sensitive and specific detection of human immunodeficiency virus type 1 in clinical speci
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Molecular Biology Today 2001. 2(2): 27-32. Vinay K. Singh and Anil Kumar Bioinformatics Sub-centr
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siRNA是小干扰rna；shRNA是小发卡rna，一段rna单链，形成茎环结构部分双链的小rna siRNA可以是天然产生也可以是人工导入的，shRNA一般都是指人工的 shRNA多是用dna载体构建，在宿主内自行转录成shRNA，再加工成像siRNA一样的小RNA，目的为了模仿。
HPLC－System－features－stackable,－modular－design
Product News Network, March 25, 2005 Finnigan™ Surveyor Plus provides PDA detection for high-throughput sample processing environments. It feature
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首先引物要跟模板紧密结合，其次引物与引物之间不能有稳定的二聚体或发夹结构存在，再次引物不能在别的非目的位点引起DNA聚合反应（即错配）。围绕这几条基本原则，设计引物需要考虑诸多因素，如引物长度（primer length）、产物长度（product length）、序列Tm值（melting t
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Comparison of lysis methodsSonicationmost efficient method of cell fractionationproblem: heat build up which can denature proteins (proportionate to l
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death Z. Xia 10/2/95The simplest way: trypan blue. Dead cells stain blueNon-fixed cells: FDA(fluorescein diacetate)-green, alive cells; P.I. (propidiu
体内表达shRNA（短发夹RNA）的设计
1.克隆到shRNA表达载体中的shRNA包括两个短反向重复序列，中间由一茎环（loop）序列分隔的，组成发夹结构，由polⅢ启动子控制。随后在连上5-6个T作为RNA聚合酶Ⅲ的转录终止子。 2.两个互补的寡核苷酸两端须带有限制性酶切位点。 3.Stratagene发现29个寡核苷酸较之原先推荐的2
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Analysis and Reconstitution of Phycobiliproteins: Methods for the Characterization of Bilin Attachment ReactionsPhycobiliproteins are a homologous fam
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实验概要Basic Methods of Culturing Drosophila实验步骤Stockkeeping1. Mechanics Most stocks can be successfully cultured by periodic mass transfer of a