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In an attempt to be concise and understandable introductory level courses and textbooks frequently present concepts that are technically correct but lead to misconceptions on the part of the student b ...

Preparation of Chromosome-sized Yeast DNA Molecules in Solid AgaroseGrow 5 ml yeast culture in YPD Broth to " stationary" (OD600 10-14)Transfer 1ml to a microfuge tube ( PGC Scientific 2.2ml tube #509 ...

These protocols should yield enough cytosol and organelles for 1-200 MT/Organelle motility assays.Solutions and Reagents Freshly removed or flash frozen rat liver PBS Homogenization Buffer Homogeniz ...

I. Useful Values 1 mg/ml tubulin = 10 µM (assuming MW of ab-tubulin heterodimer is 100000; in reality it is ~110000 but almost all tubulin labs use this convenient conversion relationship) 1 µm of a m ...

LEVEL I Figure 7.3 Isolated microsomes Figure 7.4 TEM of hepatocyte MATERIALS 1% Glutaraldehye (GTA) 1% Osmium tetroxide Epoxy or vinyl resin for TEM TEM photomicrograph of liver cells ...

Although many protocols for tubulin preparation are available the procedure described below is the simplest and highest yielding preparation I have done. The protocol calls for 3 pig brains and shoul ...

I'm trying to do a southern blot and I have a trouble the bands I get are the same bands that appear in the genomic DNA digestion my probe is specific and it works in northern blot.Thanks -PPlopez- ...

This is a quick method of getting paraformaldehyde (PFM) into solution. It is a good alternative to the traditional methods which can take hours and has the advantage that it is so quick that you are ...

　　一、概述： 　　Promega公司的SILVER SEQUENCETM DNA测序系统是一种无放射性的序列分析系统，它通过灵敏的银染方法检测凝胶中的条带。 银染提供了一种对于放射性或荧光法来说更加快速，廉价的替代方法。测序结果可以在同一天内得到；电泳完成后经90分钟就可读序，这是常规的放射性测序法做不到的。 此外 SILVER SEQUENCETM系统用未修饰的5'OH寡聚核苷酸作为引物， 减 ...

Large Scale Tubulin Preparation Tubulin is purified from bovine/porcine brain by two cycles of polymerization/depolymerization followed by removal of copurifying proteins on a phosphocellulose (PC) co ...

实验目的 将复杂的细胞分子混合物加入有机溶媒萃取以除去蛋白质及其它成分，就可以纯化DNA。一般常用酚及氯仿(phenol/chloroform)可使蛋白质变性(denaturaion)的特性来进行萃取的步骤，DNA和RNA不溶于有机溶媒中，而溶于水层。另外，分子选殖(molecular cloning)常利用洋菜胶(agarose)来分离不同大小的DNA片段或用以纯化DNA。 核酸定量可利用核酸会 ...

摘要 RNAi技术是研究基因功能的重要工具。其原理是对某个已知基因，设计诱导其沉默的dsRNA，通过合适手段导入细胞或机体使产生干涉效应的信号分子siRNA，导致基因表达水平下降或完全沉默。通过基因表型变化，鉴定该基因功能。从RNAi的研究背景和作用机制出发，对近年来利用RNAi技术研究植物基因功能的概况、诱导方法和载体作一综述。 　　关键词 基因沉默；RNAi技术；植物基因功能 　　 　　1RA ...

Mike W. Byrom Angie M. Cheng and Lance P. Forundefined Ambion Inc. 2130 Woodward Street Austin TX 78744-1832 *Corresponding author email: lford@ambion.com AbstractSmall interfering RNA (siRNA) is an extremel ...

监测手段：对脱靶效应的监测一般通过基因芯片来监测。Gene expression profiling对脱靶效应。脱靶效应的特性：转录物表达模式有序列特异性和靶特异性两种大部分脱靶效应具有siRNA序列特异性低浓度也可能产生脱靶效应，不仅仅是高浓度能产生脱靶效应的假象。脱靶效应在蛋白和mRNA水平同时考虑进行分析，要缜密分析和确定分析的时间点。特别是要确定脱靶效应是否是蛋白被knock－down的次 ...

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：对正义和反义链进行部分的phosporothioate backbone和 2’－O－methyl sugar 修饰，增加其在血清中和组织匀浆中对核酸内切酶和外切酶的抗性。 ...

虽然通过siRNA处理，整个基因的mRNA都受到影响，围绕siRNA周围的区域是受沉默影响最大的区域。就时效性研究来看，编码区是siRNA效应的最稳定区，非翻译区相对稳定性较弱。这与断裂位点和非翻译区之间的距离有关。因此引物最好选择在编码区内的任何位点，为了最大的灵敏度推荐使用围绕着siRNA位点进行设计。为了比较不同的siRNA针对不同的基因的相对沉默效果，最好在选择引物的时候用对于每个目的基因 ...

1.miRNA Map 是一个整合的数据，被开发用来存储已知miRNA 基因，假定的miRNA基因，已知的miRNA targets和假定的miRNA target.(Hsu et al 2006).2.已知的miRNA基因，来自人，小鼠，大鼠以及狗的miRNA基因，可以从miRNAase获得，试验已经证实的miRNA targets在文献中可以获得。3.假定的miroRNA precursors ...

Verifying the sequence of an shRNA hairpin is essentialsince mismatch of even one nucleotide within the target sequence can ablate knockdown . An issue that is frequently encountered in the preparatio ...

Nucleic Acids Research 2007 Vol. 35 No. 10 3339-3354S. Smit J. Widmann and R. Knighundefined Department of Chemistry and Biochemistry University of Colorado Boulder CO 80309 Understanding patterns of rRNA ev ...

用siRNA片段还是用shRNA质粒，这个要根据具体情况而定，两种方法都做是一种可行的方法，如果您经费足够。具体使用何种方法进行给药，这个具体情况具体分析，给您一个参考如下：