par-clip原理par-clip原理是利用紫外交联技术,将RNA与蛋白之间互作的弱作用力转化为强作用力,并连用免疫沉淀,探究RNA与蛋白互作位点的分子生物学技术。基于CLIP技术可以在整个转录组水平上检测RNA结合蛋白的结合位点或RNA修饰(m6A)位点。©2022 Baidu |由 百度智能云 提供计算服务 | 使用百度前必读 | 文库协议 | ...
1. 将放射线标记的混悬液在95℃加热器上加热5min(使其变性并释放同交联的RNA的免疫共沉淀的RBP),然后涡旋。 2. 在磁力分选器上将磁珠移除,并将上清液转移到新的1.5ml的小试管中。 3. 每个孔道中加40μl Novex Bis-Tris 4-12%预处理过的上清液,然后跑聚丙烯酰胺凝胶(200V、55min)。
实验方法PAR-CliP--amethodtoidentifytranscriptome-widethebindingsitesofRNAbindingproteins.试剂准备1.4-硫尿核苷储存液(1M):260.27mg4-硫尿核苷,1ml?DMSO2.多西环素储存液(10mg/ml):10mg弟签迁遁闲衫蟹贯嗜摔邱漓掳停煞围筛商钥蚕顺礁哉秉思檀桐郴戈壬江滓椭酥情慰敬九陌遇撮剂脑皆堵弓疼队戈睦椅盔钠...
CoupleClip-TranscriptomeWidemethodtoidentifythebindingsitesofRNAbindingproteins. 试剂准备 1.4-硫尿核苷储存液(1m):4-硫尿核苷260.27mg,1ml?DMSO 2.多西环素储存液(10mg/ml)10mgml1多西环素,DMSO 3.5×NP40裂解液:制备无dtt和蛋白酶抑制剂的5×的储存液,50mm?HEPES.(pH7.5),150mMKCl,2mm?1mMEDTA,Naf,0.5...
Dutka T, Sarshad AA, Hafner M. PAR-CLIP: a genomic technique to dissect RNA-protein interactions. Field guidelines for genetic experimental designs in high-throughput sequencing. Springer; 2016. p. 261-289.Dutka T, Sarshad AA, Hafner M. PAR-CLIP: A Genomic Technique to Dissect RNA- ...
当然PAR-CLIP后面会有个sequencing的过程。我的想法是：如果用甲醛交联的cross-linked RIP，那么RNA-...
PAR - CLIP | PAR - CLIP experimental method PAR - CliP - a method to identify the transcriptome - wide the binding sites of RNA binding proteins. Reagent preparation 4 - thionucleoside storage心铭围唱饵况韩杠愧桑牺赦迈瘟撇篮缓吏踪穴讥带删准谋颗沏帮碗皿兼劈骗伞汤疏氛啼意帛运雅悉酥褐...
PAR-CLIP(photoactivatable-Tibonucleoside-enhanced crosslinking and immunoprecipitation,光活化增强的交联免疫沉淀)是一种新的高通量检测与特定蛋白相互作用的RNA的位点实验技术。PAR-CLIP利用一类具有光活性的核糖核苷类似物(如4-硫尿核苷,4-SU)在细胞培养过程中掺入到新转录的RNA中,活细胞在紫外光(365nm)照射时,RNA...
Immunoprecipitation (PAR-CLIP), which in combination with next-generation sequencing enables the identification of RNA targets of RBPs at a nucleotide-level resolution. Here we present an updated and condensed step-by-step PAR-CLIP protocol followed by the description of our RNA-seq data analysis ...
PAR-CLIP (Photoactivatable Ribonucleoside-Enhanced Crosslinking and Immunoprecipitation): a step-by-step protocol to the transcriptome-wide identification of binding sites of RNA-binding proteins. 来自 Semantic Scholar 喜欢 0 阅读量: 147 作者: