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Microbial and enzyme technology: An efficient and convenient method for MiniPrep analysis of recombinant plasmids
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作者 Masahiro Sato Emi Inada +1 位作者 Issei Saitoh Yuko Matsumoto 《Journal of Biomedical Science and Engineering》 2014年第3期105-107,共3页
Minipreparation (MiniPrep) analysis is an essential step for obtaining a recombinant plasmid that carries a DNA insert containing a gene of interest. The most commonly used method for this involves cultivation of tran... Minipreparation (MiniPrep) analysis is an essential step for obtaining a recombinant plasmid that carries a DNA insert containing a gene of interest. The most commonly used method for this involves cultivation of transformed Escherichia coli (E. coli) in liquid medium, brief centrifugation for precipitation of bacterial pellets, and subsequent lysis of the pellets. This process is time-consuming and laborious, especially when the sample number is high. Here, we describe a more convenient method for MiniPrep analysis that utilizes solid medium-based cultivation of bacteria. 展开更多
关键词 AGAR PLATE E. COLI miniprep ANALYSIS Gene Engineering PLASMID
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Methodological Studies on Genomic DNA Extraction and Purification from Plant Drug Materials 被引量:12
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作者 曹晖 毕培曦 邵鹏柱 《Journal of Chinese Pharmaceutical Sciences》 CAS 1998年第3期18-25,共8页
This paper reports the rationale and methods of DNA extraction and purification from nine species of Compositae and four commercial drugs of corresponding plant Elephantopus scaber. The comparison of three methods: Cs... This paper reports the rationale and methods of DNA extraction and purification from nine species of Compositae and four commercial drugs of corresponding plant Elephantopus scaber. The comparison of three methods: CsCl gradient, CTAB/CsCl gradient and CTAB miniprep extraction by yield, purity and factors affecting PCR was carried out. In conclusion, CTAB miniprep method provides a rapid, effective, economic approach for isolating genomic DNA for Chinese drug identification by genomic fingerprints. 展开更多
关键词 DNA extraction DNA purification CTAB miniprep Genomic fingerprinting AP PCR
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海洋单细胞四爿藻基因组DNA的微量提取 被引量:8
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作者 罗立明 欧阳叶新 胡鸿钧 《武汉植物学研究》 CSCD 2003年第4期295-300,共6页
四爿藻具有坚硬的囊壳和特殊的细胞壁组成,细胞不易破碎,且含有丰富的糖蛋白,易对DNA造成污染,使其基因组DNA提取较为困难、纯度不高。研究对比传统的CTAB法与高盐低pH法提取四爿藻DNA,高盐低pH法快速经济,得到的基因组DNA纯度较高,是... 四爿藻具有坚硬的囊壳和特殊的细胞壁组成,细胞不易破碎,且含有丰富的糖蛋白,易对DNA造成污染,使其基因组DNA提取较为困难、纯度不高。研究对比传统的CTAB法与高盐低pH法提取四爿藻DNA,高盐低pH法快速经济,得到的基因组DNA纯度较高,是一种行之有效的四爿藻基因组DNA提取方法。该法通过改变抽提介质,提高细胞破碎效率,减少抽提次数,有效去除污染,提取的基因组DNA不仅可以成功进行nrDNA转录间隔区(ITS)扩增,而且扩增产物适合于进行测序分析,这为其它淡水和海洋单细胞及多细胞藻类基因组DNA的提取也提供了借鉴。 展开更多
关键词 四爿藻 基因组DNA提取 高盐低pH法 CTAB法
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结直肠癌组织中K-ras基因的检测及其意义 被引量:8
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作者 徐向明 张国良 +2 位作者 李敏伟 王丹阳 林建江 《中华医学杂志》 CAS CSCD 北大核心 2010年第18期1268-1271,共4页
目的 观察结直肠癌组织中K-ras基因的表达情况,并分析其与临床病理学的相互关系,为临床筛选出能从生物靶向治疗中获益的人群.方法 对208例结直肠癌组织标本,使用聚合酶链式反应(PCR)技术扩增目的基因(包含12和13密码子),并对扩增产... 目的 观察结直肠癌组织中K-ras基因的表达情况,并分析其与临床病理学的相互关系,为临床筛选出能从生物靶向治疗中获益的人群.方法 对208例结直肠癌组织标本,使用聚合酶链式反应(PCR)技术扩增目的基因(包含12和13密码子),并对扩增产物进行Sanger法测序,进行K-ras基因的检测,结合相关的临床病理学资料,分析K-ras基因的表达状况同临床病理学的关系.结果 208例结直肠癌组织中K-ras基因12或13密码子突变为91例,突变率为43.8%,突变率在男女性别之间、肿瘤位置、组织分化类型和Duke's分期间差异无统计学意义.结论 对结直肠癌组织K-ras基因的表达状况进行检测有助于临床筛选出针对抗EGFR靶向治疗药物有效的结直肠癌患者. 展开更多
关键词 结直肠肿瘤 基因 ras 靶向治疗
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DNA purification and gene typing: Based on multifunc- tional nanobeads 被引量:1
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作者 XIExin ZHANGXu +4 位作者 GAOHuafang ZHANGHuan CHENDepu CHENGJing FEIWeiyang 《Chinese Science Bulletin》 SCIE EI CAS 2004年第9期886-889,共4页
In this report, a universal protocol for extract-ing genomic DNA from whole blood, saliva, and bacterial culture by using magnetic nanobeads as solid-phase absor-bents was presented. The enrichment of target cells and... In this report, a universal protocol for extract-ing genomic DNA from whole blood, saliva, and bacterial culture by using magnetic nanobeads as solid-phase absor-bents was presented. The enrichment of target cells and ad-sorption of DNA have been functionally integrated onto the surfaces of the carboxyl-modified magnetic nano-beads, and the DNA segments bound on the surface of the beads can be directly used as PCR templates to amplify a target gene. The PCR products were applied to an oligonucleotide array to perform gene typing. The protocol proves to be simple, rapid, biologically and chemically nonhazardous, and promising for the microfabrication of DNA preparation chip. 展开更多
关键词 多功能纳米床 DNA纯化 染色体组DNA 微片 PCR 固相吸收 显微完全分析系统 诊断 R44
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