小分子干扰RNA (Small interfering RNA ，siRNA ) 是一种非常有效的工具，能够在包括哺乳动物细胞在内的多个体系中抑制特定基因的表达，从而研究某个基因的功能或者是相关信息。但是这种强有力的方法的难题之一是需要设计，合成siRNA s 和验证其效果，从而找到最有效的siRNA s 。
1.NCBIshRNA资源库 http://cgap.nci.nih.gov/RNAi About RNAi on CGAPThe NCBI is part of the consortium supporting the preparation of human and mouse libraries containing RNAi constructs that target cancer-r ...
差异基因表达的研究受到了广泛的关注，常用的技术有DD-PCR；GENE-FISHING；GENE CHIP等。简单介绍如下： DDRT―PCR技术即mRNA差异显示聚合酶链式反应技术，此技术是以PCR技术和聚丙烯凝胶电泳技术为基础，结合银染或放射性自显影等显色技术，能快速有效地鉴定并克隆两个或多个平行材料之间的差异表达基因。DDRT―PCR技术的基本过程如下：①提取两组平行材料中的总RNA或m ...
编者按：不同生物中RNA 结构和功能多样性与生命多样性的关系等问题是当前生命科学重大基础研究的前沿，特别随着基因组、蛋白质组计划相继提出后，RNA 组学研究已经成为分子生物学研究的新的生长点。本期RNA 专题由王恩多院士牵头，共组织了八篇与RNA 研究相关的高质量文章。在此，向王恩多院士及各位专家对本刊的大力支持表示衷心的感谢！张筱晨， 周 惠， 屈良鹄~A中山大学生物工程研究中心，广州51027 ...
刘默芳，王恩多(中国科学院上海生命科学研究院生物化学与细胞生物学研究所，分子生物学国家重点实验室，上海200031)摘 要：小分子调控RNA，包括siRNA (small interfering RNA)、miRNA (microRNA)和piRNA (piwi interacting RNA)、hsRNA (heterochromatin associated small RNA)等，是当前生 ...
1.相似性序列发生Cross－react引起脱靶效应。2.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。3.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。4.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。
Overview With 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 ...
RNA extraction buffer:0.1M NaCl2% SDS50mM Tris/HCl (pH9)10mM EDTA20mM %26szlig;-mercaptoethanol1. Grind leaf tissue on liquid N in mortar and pestle.2. Transfer the ground tissue to a 10ml conical bot ...
Steve Hahn. last modified Sat Oct 17 1998Mix the following in an 0.5 ml eppendorf tube:10-20 micrograms total yeast RNA10 microliters hybridization mixH2O to bring the final volume to 25 microliters40 ...
人、小鼠和大鼠的多种组织来源的高质量mRNA经电泳分离后预转于尼龙膜上预制的即用型Northern杂交膜，免去RNA电泳操作过程可检测范围：0.5-10kb可重复使用提供ExpressHybTM快速杂交液样品可靠的优良品质从1991年始，公司就向广大的生命科学研究者提供高质量的预制杂交膜。而MTN™膜更是在国际著名的学术期刊上有将近3000篇文献引用。严谨的制作过程1.由人、小鼠和大鼠的组织经Ol ...
siRNA Database Searchable database of Silencer ™ Validated and Pre-designed siRNAs to 34000 human mouse and rat targets. All siRNAs in the database have been designed for maximum potency and specifici ...
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 ...
In vitro transcription with yeast nuclear extract Steve HahnWear gloves throughout use RNAse free solutions (either autoclaved or sterile filtered) and clean bench and pipetmen with 95% ethanol before ...
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 ...
Run-on transcription monitors the regulation of transcription in isolated nuclei.A. Preparation of Nuclei - (do everything at 0℃ to 4℃ in 50 ml tubes)1. Pellet between 30 and 300 million cells at 1500 ...
Polymerase III in vitro Transcription Steve HahnFor the following reactions use appropriate shielding and dispose of radioactive waste properly!A 20 microliter transcription reaction contains:4.0 μ ...
1 ml linearized DNA template pBS-SK- ERD (X-E) (0.1-0.5 mg)7.8 ml DDW5 ml 5X transcription buffer ( stratagene )1 ml 750 mM DTT1 ml Rnasin (RNase inhibitor; 15U/ml)1 ml 10 mM ATP (pH 7)1 ml 10 mM CTP ...
The following protocol is for MEGAscript II Kit (Ambion). 1. PCR amplify the DNA template. The 5'-end of the template should contain the minimum promoter sequenences of T7 or Sp6 or T3.A 5-primer with ...
Transcription of cRNA probeReagents/SolutionsDNA (PCR or plasmid) with bacteriophage RNA polymerase promoter in suitable orientation (see note 1 ) ~1µg/µl in TE buffer 10mM MgCl2 5x Transcription Buff ...
T℃OS-M6 cells grow in 6-well plates 2ml media total T℃V1PD cells grow in 100mm plates.1: Aspirate off the media from the cells and was the cell monolayers with 2x5ml (To 100mm dish) or 2x1ml (to 6-wel ...
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