酵母遗传学技术-实验方法

实验频道正文用户登录新用户注册酵母遗传学技术作者：佚名实验频道来源：中华基因网点击数：更新时间：2004-6-8 Genome-wide Gene Expression Analysis (Richard Young Research Group,Whitehead Institute for Biomedical Research)A genoe-wide gene expression analysis using high-density oligonucleotide arrays Restriction Digests of High Molecular Weight Yeast DNA (Donis Keller Lab)Restriction digests of YACs for mapping using rare cutting enzymes or more conventional restriction endonuclease digestion. Preparation of Yease Cell for ImmunoEletron Microscopy (Jon Mulholland, Botstein Lab) Yeast Immunofluorescence with Methanol/Acetone Dehydration (Botstein Lab) Yeast Immunofluorescence (without methanol) (D. Amberg, Botstein Lab) Yeast Rhodamine-Phalloidin/Calcofluor Staining (D. Amberg, Botstein Lab) Yeast Cell Cycle by Flow Cytometry (Salk Institute, Flow Cytometry Laboratory) Vectorette PCR of Yeast DNA (Botstein Lab) Yeast Actin Capture Assay (D. Amberg) Yeast Preparation for FACS Analysis (Gottschling Lab) Replication Timing by Density Transfer (Fangman/Brewer lab) Replication Timing by Comparative Hybridization (Fangman/Brewer lab) Replication Timing Using Transient Hemimethylation (Fangman/Brewer lab) ß-GAL Filter Assay (Breeden Lab) Bgal filter assay (Herskowitz Lab) Bgal plate overlay assay (Herskowitz Lab)This assay is semiquantitative. Cells can even be recovered from underneath the overlaid agar. The method could be used for any enzyme with a colorimetric assay. Bgal liquid assay (Herskowitz Lab) Pheromone Halo Assay (Dohlman Lab)This is a bio-assay that measures the responsiveness of cells to a factor pheromone. The assay is easy to conduct and the results are usually unambiguous and highly reproducible. Liquid ß-gal Assays (Dohlman Lab)This protocol is used to quantitatively measure the pheromone response over a range of pheromone concentrations. ß-gal Lift Protocol (Dohlman Lab) This method is used to screen for and examine mutants with altered pheromone responses. Handling and using insertion libraries (YGAC) Curing strains of endogenous 2-micron mTn-lacZ/LEU2 insertion library mTn-3xHA/lacZ insertion library mTn-3xHA/GFP insertion library Yeast Gene Mutagenesis Protocols for shuttle mutagenesis/epitope-tagging of a yeast gene with mTn-lacZ/LEU2 (Yale Yeast Analysis Center)mTn-lacZ/leu2 can easily be inserted at mutiple sites in a given gene. The mutagenized DNA is then transformed into yeast, where it replaces the chromosomal locus by homologous recombination. The transposon insertions create a pool of insertion/disruption alleles. Insertions that generate in-frame fusion of the coding region to lacZ can be used to monitor and quantify gene expression, via assays for beta-gal activity. The fusion protein can also be immunodetected using antibodies directed against beta-gal. Protocols for shuttle mutagenesis/epitope-tagging of a yeast gene with mTn-3xHA/lacZ (Yale Yeast Analysis Center)Same protocol as above Mutagenizing a yeast gene with an mTn (YGAC) Non-technical overview Using mTn-lacZ/LEU2 Using mTn-3xHA/lacZ Using mTn-4xHA/lacZ Using mTn-3xHA/GFP UV Mutagenesis (Corbett Lab) EMS Mutagenesis (Herskowitz Lab) EMS mutagenesis of yeast (Hahn Lab) Generating a Temperature Sensitive Allele (Herskowitz Lab) Confirmation PCR (Saccharomyces Genome Deletion Project)To study the genetics of yeast, yeast gene ORF is deleted by PCR based strategy to generate mutated strain. Confirmation PCR is used to verify if the yeast ORF deletion is successful Introduction/General strategyConfirmation PCR primer designPrimer organization in the 96-well platesOligonucleotide concentrationsQuality control proceduresSingle tube PCR protocol96-well format protocol 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：酵母人工染色体下一篇实验频道：酵母双杂交系统【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细胞培养的微生物污染排除质粒的酵母直接转化酵母遗传学方法12：酵母菌落酵母遗传学方法11：PCR介导基酵母遗传学方法10：固定化细酵母遗传学方法9：酵母免疫荧酵母遗传学方法8：酵母活体染酵母遗传学方法7：随机孢子分酵母遗传学方法6：羧肽酶Y的酵母遗传学方法5：酵母β－半 · 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
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