Gene analysis techniques最新文献

筛选
英文 中文
Cytogenetic methodologies for gene mapping and comparative analyses in mammalian cell culture systems 哺乳动物细胞培养系统中基因定位和比较分析的细胞遗传学方法
Gene analysis techniques Pub Date : 1987-07-01 DOI: 10.1016/0735-0651(87)90021-5
William S. Modi , William G. Nash , Anna C. Ferrari , Stephen J. O'Brien
{"title":"Cytogenetic methodologies for gene mapping and comparative analyses in mammalian cell culture systems","authors":"William S. Modi ,&nbsp;William G. Nash ,&nbsp;Anna C. Ferrari ,&nbsp;Stephen J. O'Brien","doi":"10.1016/0735-0651(87)90021-5","DOIUrl":"10.1016/0735-0651(87)90021-5","url":null,"abstract":"<div><p>Presented here are the detailed methods employed in our laboratory for gene mapping and cytogenetic analyses in human beings, in the domestic cat, and in other mammalian species. Included in the procedures are: 1) establishment of primary fibroblast and lymphoid cell cultures; 2) heterologous cell fusion for production of rapidly proliferating cell hybrids; 3) cellular transformation of primary fibroblasts using an oncogenic retrovirus; 4) cell synchronization for high-resolution banding of prometaphase chromosomes; 5) chromosome-banding procedures, including G-banding, alkaline G-11, and Q-banding; and 6) in situ hybridization of radiolabeled molecular clones to metaphase chromosomes for regional gene localization.</p></div>","PeriodicalId":77714,"journal":{"name":"Gene analysis techniques","volume":"4 4","pages":"Pages 75-85"},"PeriodicalIF":0.0,"publicationDate":"1987-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0735-0651(87)90021-5","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"14629185","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 48
Electric field-mediated gene transfer (electroporation) into mouse friend and human K562 erythroleukemic cells 电场介导的基因转移(电穿孔)到小鼠朋友和人K562红白血病细胞
Gene analysis techniques Pub Date : 1987-05-01 DOI: 10.1016/0735-0651(87)90018-5
Demetrios A. Spandidos
{"title":"Electric field-mediated gene transfer (electroporation) into mouse friend and human K562 erythroleukemic cells","authors":"Demetrios A. Spandidos","doi":"10.1016/0735-0651(87)90018-5","DOIUrl":"10.1016/0735-0651(87)90018-5","url":null,"abstract":"<div><p>Electroporation, the technique of electric field mediated gene transfer, was evaluated as a means of introducing and expressing genes into mouse Friend and human K562 erythroleukemic cells. Long-term (stable) gene expression in both Friend and K562 cells was measured using the recombinant plasmid Homer 6, which carries the aminoglycoside phosphotransferase (<em>aph</em>) gene as a selectable marker under the transcriptional control of the Moloney murine sarcoma virus long terminal repeat promoter/enhancer sequences. Parameters such as the DNA concentration, the initial field strength, the concentration of recipient cells, and the preselection expression time were examined to obtain optimal transfection frequencies. Short-term (transient) expression was also examined using the plasmid pLW4, which carries the chloramphenicol acetyltransferase gene under the transcriptional control of herpes simplex virus immediate early 5 gene promoter/enhancer sequences. Conditions that gave maximal stable transformation frequency were similar to those giving highest transient gene expression in the mouse and human erythroleukemic cell lines. Under optimal conditions, electroporation gave about ten times higher transfection frequencies and levels of transient expression for both types of cells when compared with the calcium phosphate technique. Because both Friend and K562 cells can be induced to differentiate in vitro, measurement of transient or stable expression levels for genes introduced into these cells may prove to be useful in the study of developmental regulation of genes from the erythroid pathway.</p></div>","PeriodicalId":77714,"journal":{"name":"Gene analysis techniques","volume":"4 3","pages":"Pages 50-56"},"PeriodicalIF":0.0,"publicationDate":"1987-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0735-0651(87)90018-5","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"14631466","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 22
Simultaneous sequencing of RNA by solid-phase chemical degradation using DE 81 anion-exchange paper 用de81阴离子交换纸固相化学降解法同时测序RNA
Gene analysis techniques Pub Date : 1987-05-01 DOI: 10.1016/0735-0651(87)90019-7
Burckhard Gross, André Rosenthal
{"title":"Simultaneous sequencing of RNA by solid-phase chemical degradation using DE 81 anion-exchange paper","authors":"Burckhard Gross,&nbsp;André Rosenthal","doi":"10.1016/0735-0651(87)90019-7","DOIUrl":"10.1016/0735-0651(87)90019-7","url":null,"abstract":"<div><p>A solid-phase chemical degradation method for simultaneous sequencing of RNA and RNA fragments has been developed using Whatman DE 81 anion-exchange paper as the support. The approach involves the following operations: (1) immobilization of the 3′-end labeled RNAs or RNA fragments on DE 81 paper; (2) washing; (3) modification reactions; (4) washing; (5) sorting of the paper segments; (6) aniline reaction; (7) lyophilization; (8) desorption of the RNA by TEAB; and (9) lyophilization.</p></div>","PeriodicalId":77714,"journal":{"name":"Gene analysis techniques","volume":"4 3","pages":"Pages 57-61"},"PeriodicalIF":0.0,"publicationDate":"1987-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0735-0651(87)90019-7","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"13612286","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 4
Simultaneous isolation of high molecular weight RNA and DNA from limited amounts of tissues and cells 同时从有限数量的组织和细胞中分离高分子量RNA和DNA
Gene analysis techniques Pub Date : 1987-05-01 DOI: 10.1016/0735-0651(87)90017-3
Eckart Meese, Nikolaus Blin
{"title":"Simultaneous isolation of high molecular weight RNA and DNA from limited amounts of tissues and cells","authors":"Eckart Meese,&nbsp;Nikolaus Blin","doi":"10.1016/0735-0651(87)90017-3","DOIUrl":"10.1016/0735-0651(87)90017-3","url":null,"abstract":"<div><p>A simple method is described for the simultaneous isolation of both DNA and RNA from tissues and cultured cells obtainable in limited quantities only. The method is based on a suitable combination of steps designed for preparations of high molecular weight nucleic acids in cases when restricted amounts of tissues like small-sized and unique biopsies of tumors are available for studies of gene organization and expression. Using this protocol, undegraded total RNA suitable for Northern blot analysis and high molecular weight DNA for Southern blots was obtained from various sources (mammary and colon carcinomas, meningiomas, colonic and placental tissue, and several cell cultures).</p></div>","PeriodicalId":77714,"journal":{"name":"Gene analysis techniques","volume":"4 3","pages":"Pages 45-49"},"PeriodicalIF":0.0,"publicationDate":"1987-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0735-0651(87)90017-3","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"13612284","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 25
Preparation of large numbers of plasmid DNA samples in microtiter plates by the alkaline lysis method 用碱性裂解法在微滴板上制备大量质粒DNA样品
Gene analysis techniques Pub Date : 1987-05-01 DOI: 10.1016/0735-0651(87)90016-1
Toby J. Gibson, John E. Sulston
{"title":"Preparation of large numbers of plasmid DNA samples in microtiter plates by the alkaline lysis method","authors":"Toby J. Gibson,&nbsp;John E. Sulston","doi":"10.1016/0735-0651(87)90016-1","DOIUrl":"10.1016/0735-0651(87)90016-1","url":null,"abstract":"<div><p>A protocol is described for the growth and preparation of plasmid DNAs from small culture volumes (250 μl) and utilizing standard 96-well plates. Several hundred plasmids can be prepared simultaneously, yielding sufficient DNA for subsequent analysis by restriction digestion and gel electrophoresis. This protocol may be useful for rapid screening of clones arising in recombinant DNA work such as site-directed mutagenesis, oligonucleotide cassette cloning, deletion analysis, etc. The technique was initially developed to meet our requirement to provide large numbers of cosmid DNAs for restriction enzyme fingerprint analyses in genome mapping projects.</p></div>","PeriodicalId":77714,"journal":{"name":"Gene analysis techniques","volume":"4 3","pages":"Pages 41-44"},"PeriodicalIF":0.0,"publicationDate":"1987-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0735-0651(87)90016-1","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"14462108","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 23
Hybridization of Oligo(dT) to RNA on nitrocellulose 寡核苷酸(dT)与RNA在硝化纤维素上的杂交
Gene analysis techniques Pub Date : 1987-03-01 DOI: 10.1016/0735-0651(87)90013-6
Calvin B. Harley
{"title":"Hybridization of Oligo(dT) to RNA on nitrocellulose","authors":"Calvin B. Harley","doi":"10.1016/0735-0651(87)90013-6","DOIUrl":"10.1016/0735-0651(87)90013-6","url":null,"abstract":"<div><p>Quantitation of mRNA immobilized on nitrocellulose filters is an essential aspect of some studies in molecular biology. Hybridization of oligo(dT)<sub>18</sub> to the poly(A) tails of mRNA can be used to measure filter-bound mRNA and thus provides a basis for comparing abundance of specific mRNAs. Hybridization rate of <sup>32</sup>P-labeled oligo(dT)<sub>18</sub> in 0.75 M NaCl, 75 mM sodium citrate, pH 7 (5 × SSC) to immobilized RNA was maximal at 25°C. Filters were fully hybridized under these conditions within 1 hr when the oligo(dT)<sub>18</sub> concentration was 10 pmol/ml or higher. Salt dependence of the dissociation temperature (T<sub>d</sub>) of oligo(dT)<sub>18</sub>:RNA duplex on filters was described by the equation T<sub>d</sub> = 42 − 20log<sub>10</sub>[molar Na<sup>+</sup>] (°C). With stringent washing of the duplex (four 5-min washes in 2 × SSC at room temperature), oligo(dT)<sub>18</sub> gave no signal with plasmid DNA, rRNA, or tRNA. We have found that olig(dT)<sub>18</sub> can be used to normalize signal strengths rapidly and conveniently from total or oligo(dT)-selected eukaryotic RNA.</p></div>","PeriodicalId":77714,"journal":{"name":"Gene analysis techniques","volume":"4 2","pages":"Pages 17-22"},"PeriodicalIF":0.0,"publicationDate":"1987-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0735-0651(87)90013-6","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"13612280","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 171
Sequence analysis of proteins 蛋白质序列分析
Gene analysis techniques Pub Date : 1987-03-01 DOI: 10.1016/0735-0651(87)90015-X
Devjani Chatterjee, Jacob V. Maizel Jr.
{"title":"Sequence analysis of proteins","authors":"Devjani Chatterjee,&nbsp;Jacob V. Maizel Jr.","doi":"10.1016/0735-0651(87)90015-X","DOIUrl":"10.1016/0735-0651(87)90015-X","url":null,"abstract":"<div><p>There has been a rapid increase in the number of available protein sequences derived from gene-sequence information. Computer-based sequence analysis of proteins is gaining in importance as an analytical tool. With the help of these analyses such sequences may be characterized and some insights gained into their probable role in the system. The principles involved in computer-based sequence analysis and some of the methods are discussed.</p></div>","PeriodicalId":77714,"journal":{"name":"Gene analysis techniques","volume":"4 2","pages":"Pages 27-40"},"PeriodicalIF":0.0,"publicationDate":"1987-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0735-0651(87)90015-X","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"14629183","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 1
Storage of spheroplasts at −70°C for transfection with φX174 RFDNA 球质体保存于- 70°C,转染φX174 RFDNA
Gene analysis techniques Pub Date : 1987-03-01 DOI: 10.1016/0735-0651(87)90014-8
H.V. Malling , M.C. Fater , J.G. Burkhart , S.C. Hardies , C.A. Hutchison III , M.H. Edgell
{"title":"Storage of spheroplasts at −70°C for transfection with φX174 RFDNA","authors":"H.V. Malling ,&nbsp;M.C. Fater ,&nbsp;J.G. Burkhart ,&nbsp;S.C. Hardies ,&nbsp;C.A. Hutchison III ,&nbsp;M.H. Edgell","doi":"10.1016/0735-0651(87)90014-8","DOIUrl":"https://doi.org/10.1016/0735-0651(87)90014-8","url":null,"abstract":"<div><p>A method was developed for preparation of spheroplasts used for transfection with φX174 RF DNA. This method had a high-level competence and retained the competence for up to one year of storage in 7% DMSO at −70°C.</p></div>","PeriodicalId":77714,"journal":{"name":"Gene analysis techniques","volume":"4 2","pages":"Pages 23-26"},"PeriodicalIF":0.0,"publicationDate":"1987-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0735-0651(87)90014-8","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71867485","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 4
Efficient purification of mouse monoclonal antibodies from ascites fluid by medium-performance anion exchange chromatography 中效阴离子交换色谱法纯化小鼠腹水单克隆抗体
Gene analysis techniques Pub Date : 1987-01-01 DOI: 10.1016/0735-0651(87)90005-7
Michael E Annunziato, Dante J Marciani
{"title":"Efficient purification of mouse monoclonal antibodies from ascites fluid by medium-performance anion exchange chromatography","authors":"Michael E Annunziato,&nbsp;Dante J Marciani","doi":"10.1016/0735-0651(87)90005-7","DOIUrl":"10.1016/0735-0651(87)90005-7","url":null,"abstract":"<div><p>Medium-performance anion-exchange chromatography was applied to the purification of murine IgG class monoclonal antibodies from ascites fluid. The separations were performed under mild conditions at pH 8 using relatively low sodium-chloride concentrations. Recoveries for monoclonal antibodies of subclasses IgG1, IgG2a, and IgG2b were about 90%. The IgG preparations were free of other ascites fluid proteins.</p></div>","PeriodicalId":77714,"journal":{"name":"Gene analysis techniques","volume":"4 1","pages":"Pages 1-4"},"PeriodicalIF":0.0,"publicationDate":"1987-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0735-0651(87)90005-7","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"14629177","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 4
Improvements in production of cDNA libraries in λgtll detection of ligation inhibitor and separation of cDNA from free linkers λgtll连接抑制剂检测及游离连接体分离cDNA文库的改进
Gene analysis techniques Pub Date : 1987-01-01 DOI: 10.1016/0735-0651(87)90008-2
Michael E. Pape, Ki-han Kim
{"title":"Improvements in production of cDNA libraries in λgtll detection of ligation inhibitor and separation of cDNA from free linkers","authors":"Michael E. Pape,&nbsp;Ki-han Kim","doi":"10.1016/0735-0651(87)90008-2","DOIUrl":"https://doi.org/10.1016/0735-0651(87)90008-2","url":null,"abstract":"<div><p>A ligation assay was designed to determine if a ligation inhibitor is present in a Bio-Gel A-50-m column, the traditional tool used for separation of cDNA from free linkers during the λgtll cloning procedure. A new and simple electrophoresis method is also presented for performing this separation, which is rapid, efficient, and reduces the chance of carrying along a ligation inhibitor.</p></div>","PeriodicalId":77714,"journal":{"name":"Gene analysis techniques","volume":"4 1","pages":"Pages 14-16"},"PeriodicalIF":0.0,"publicationDate":"1987-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0735-0651(87)90008-2","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71868095","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
0
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
相关产品
×
本文献相关产品
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信