RNA标记-实验方法

实验频道正文用户登录新用户注册 RNA标记作者：佚名实验频道来源：中华基因网点击数：更新时间：2004-6-8 RNA labeling (Amersham Pharmacia)For the generation of radiolabelled, single stranded RNA for use as hybridization probes 32P-pCp 3 End-labeling RNA (Jim Brown Lab)This procedure includes two methods for the synthesis of 32P-pCp, because of the enormous cost of the commercial product (about $1 per uCi). The compound is easy to synthesis, & is fresher than the commercial product. RNA end-labeling (gopher://iubio.bio.indiana.edu)RNA can be 5 or 3 end-labeled. 3 end-labeling is carried out by RNA ligase, which ligates a single labeled nucleotide to the 3-OH end of the RNA. 5 end-labeling is carried out exactly as for DNA (see that procedure in M&M as well), unless the 5 ends are capped; in that case, the RNA must first be de-capped with tobacco acid pyrophosphatase (TAP).and 5 labeling Digoxigenin-UTP RNA Labeling (Frank Bottone Jr.) Preparing 5 end-labeled alkaline hydrolyzed RNA (Jim Brown Lab)The best way to obtain high specific activity labeled rRNAs is to hydrolyze the RNA by alkaline treatment at high temperature, followed by 5 end-labeling with PNK & gamma-32P-ATP. This way, you get many 32Ps incorporated into each rRNA molecule. This is also useful in heterologous probings, because the similarities of some of the RNA fragments will be much higher than the overall similarity for the heterologous rRNAs. Sephadex G-50 Spin Column (gopher://iubio.bio.indiana.edu)For purifying labeled probe 处理 SSI 文件时出错处理 SSI 文件时出错【评论】【进论坛看】【转入博客】实验频道录入：生物编辑责任编辑：生物编辑上一篇实验频道：差异表达下一篇实验频道：原位转录【发表评论】【加入收藏】【告诉好友】【打印此文】【关闭窗口】相关文章最新热门推荐文章病毒DNA和RNA的简易纯化技术哺乳动物细胞中进行RNA干扰:哺乳动物细胞总RNA的分离RNA酶活性的控制mRNA的分离2%多聚甲醛固定剂配方二则细胞学技术细胞培养的一般技术血小板的活化、荧光染色与流血小板活化检测方法 · 免疫细胞表面抗原分子CD家族对照表· 所有的看家基因(housekeeping gene· 生物发光与化学发光专题· 生物发光和化学发光在生物技术中的· 重组DNA的分离、克隆与测序实验手册· RNAi和基因沉默的历史回顾· RNAi原理图解· RNAi术语表· siRNA设计指南· Methods for DNA sequencing · 通用电气医疗集团简介· 生物发光与化学发光专题· 生物发光和化学发光在生物技术中的· RNA干扰技术获得新突破· 蛋白质特性与分离纯化技术的选择· PCR实验指导与常见问题分析· RNAi原理FLASH演示· Extrachromosomal elements, plasm· Promoter analysis by saturation · 重组DNA的分离、克隆与测序实验手册文章评论（只显示最新10条。评论内容只代表网友观点，与本站立场无关！）生物谷 | 网站介绍 | 网站地图 | 广告服务 | 帮助关于我们 | 广告服务 | 战略伙伴 | 留言| 帮助信息| 联系方式 | 客户投诉 | 友情链接 | 网站地图 | 网站声明 | Bioon English 上海北岸信息技术有限公司公司地址：上海龙吴路51号嘉源商务中心1#211室(200232) 生物谷网站 bioon.com © Copyright 2001 - 2006. All rights reserved. 备案号：沪ICP备05022939号
< 1 > < 2 >

设为首页 | 加入收藏 | 广告服务 | 友情链接 | 版权申明