TheTRCshRNADesignMethodsandRules

OverviewWe 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 rules, including those derived from the siRNA literature, our cloning scheme, constraints on the synthesis of the oligonucleotides and others. In applying the algorithm, our aim is to achieve a balance of two competing go......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 Specificity We are forced to balance the prediction of knockdown efficiency against the desire to minimize intera

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

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

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

RNAi片段siRNA设计原则

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

Northern-protocol(RULES-FOR-RNA-WORK)

1.         Wear gloves at all time including filling pipet tip in racks, filling jars with Eppendorf tubes, and weighing chemicals to prepare solution

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

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

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

shrna引物退火原理

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

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

TaqMan-Primer-and-Probe-Design

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

PCR-Primer-Design(一)

Molecular Biology Today 2001. 2(2): 27-32.                                                    Vinay K. Singh and Anil Kumar   Bioinformatics Sub-centr

PCR-Primer-Design(二)

  Terminal Nucleotides Make a Difference   Both the terminals of the primer are of vital importance for a successful amplification. The 3'-end

PCR-Primer-Design(三)

  References   Albert, J., and Fenyo, E.M. 1990. Simple, sensitive and specific detection of human immunodeficiency virus type 1 in clinical speci

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

Design-Weighing-Processes-with-Quality-in-Mind

June 15, 2016 - Greifensee, Switzerland With QbD-Quality by Design-weighing processes help ensure excellent consistency in final products By Tobias

PCR-PRIMER-DESIGN-AND-REACTION-OPTIMISATION

ContentsFactors Affecting the PCR  Nested Primer PCRPrimer LengthDegenerate PrimersElongation Temperature and TimeReaction BufferCycle NumberDenaturin

MethPrimer--Design-Primers-for-Methylation-PCRs

Welcome to MethPrimer MethPrimer is a program for designing bisulfite-conversion-based Methylation PCR Primer. Currently, it can design primers for tw

miRNA,siRNA,shRNA有什么区别

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

引物设计(Primer-Design)的原则

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

Comparison-of-lysis-methods

Comparison of lysis methods Sonication most efficient method of cell fractionation problem: heat build up which can denature proteins (proport

Staining-Methods-for-cell

death Z. Xia 10/2/95 The simplest way: trypan blue.  Dead cells stain blue Non-fixed cells: FDA(fluorescein diacetate)-green, alive cells; 

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

miRNA、siRNA、dsRNA与shRNA-表示什么意思?

siRNA,shRNA,dsRNA,miRNA间的区别RNA干扰(RNAi),即用20多个核苷酸组成的短的双链RNA(siRNA)代替传统反义核酸进行转录后基因沉默,已经迅速而广泛地应用到基因功能,基因表达调控机制研究等热门领域,并为基因治疗开辟了新的途径。近两年来,这方面的科学论文及报道爆炸性增长

General-Laboratory-Methods-for-Tetrapyrroles

There are thousands of porphyrins and metalloporphyrins, and hundreds of new derivatives appear each year. This variety arises because the cyclic

Plant-Genome-Annotation-Methods

Annotation of plant genomic sequences can be separated into structural and functional annotation. Structural annotation is the foundation of all