利用RAPD技术进行植物性状标记及辅助选择

近等基因系、混合分离群体法是ＲＡＰＤ 标记的主要策略。目前,ＲＡＰＤ标记广泛用于抗线虫、抗病、雄性不育等辅助选择的研究中,取得了可喜的成绩。由于遗传距离的不同,使ＲＡＰＤ 标记具有基因型的差异。寻找无重组的ＲＡＰＤ 标记或将ＲＡＰＤ标记转化为ＲＦＬＰ标记,可以解决这一问题。随着连锁程度的降低选择效率也随着降低。相斥相的ＲＡＰＤ标注可提高选择效率将ＲＡＰＤ标记转化为ＳＣＡＲｓ、ＡＰＳＰｓ 标记,可以解决ＲＡＰＤ 标记稳定性差的问题。来源于ＲＡＰＤ 标记的ＳＣＡＲｓ 标记将在辅助选择中发挥巨大的作用。     

大鼠RAPD标记的观察

采用随机扩增多态ＤＮＡ（ＲＡＰＤ）技术,分析ＳＤ和Ｗｉｓｔａｒ二种大鼠的基因多态性,探讨用ＲＡＰＤ标记鉴别二种大鼠及其血标本实验中的认证,结果表明,二种大鼠表现出了各自不同的多态性ＲＡＰＤ标记,作为大鼠的分子标记,可在基因水平区别二种大鼠,故认为是一种大鼠研究的分子依据。     

RAPD技术在系统与进化植物学中的应用

随着ＲＡＰＤ技术的发展,这一在ＤＮＡ分子水平上检测遗传多样性的方法被广泛用于植物系统与进化的研究之中,利用ＲＡＰＤ的分析结果,不仅可以探讨 物种间的亲缘演化关系,检测种内的遗传分化,而且还可以为属、种的分类提供强有力的证据。事实证明,ＲＡＰＤ在系统与进化植物学的研究中具有重要的参考价值,并已取得了可喜的成果。     

植物标本采集标签计算机印制数据系统

植物标本采集标签计算机印制数据系统陈涛,陈都,吴德邻（中国科学院华南植物研究所,广州５１０６５０）ＡＮＩＮＴＲＯＤＵＣＴＩＯＮＴＯＴＨＥＤＡＴＡＳＹＳＴＥＭＦＯＲＣＯＭＰＵＴＥＲ－ＡＩＤＥＤＬＡＢＥＬＰＲＥＰＡＲＡＴＩＯＮＯＦＳＰＥＣＩＭＥＮＣＯＬＬ...     

植物RAPD标记的可靠性研究

随着ＲＡＰＤ技术应用范围越来越广,ＲＡＰＤ的优缺点也不断反映出来。相左实验结果的出现,对ＲＡＰＤ标记可靠性提出了置疑。本文从重复性、序列同源性和系统学价值三个方面,就ＲＡＰＤ标记的可靠性研究的现状进行了综述。并就怎样提高ＲＡＰＤ标记的可靠性进行了探讨     

种质资源鉴定的一种分子遗传技术标准：随机扩增多态性DNA（RAPD）

ＲＡＰＤ（随机扩增多态性ＤＮＡ）变异越来越多地被用于种质样品的鉴定。由于在某些条件下,ＲＡＰＤ结果重性较差,限制这一技术的应用。本文综合了一些文献中引起片段变异的主要原因,分析了减少这些不需要变异的方法。由此提出了一个含有全部描述符的项目表,它包括ＲＡＰＤ必不可少的条件和再现植物材料ＲＡＰＤ结构的必须核对的程序。此标准程序和描述形式有促进类似材料在不同实验室所做结果进行比较,而且允许ＲＡＰＤ结果作     

中国蝮属蛇类的RAPD分析

用ＲＡＰＤ技术研究了我国５种蝮属蛇类（中介蝮，蛇岛蝮，短尾蝮，高原蝮，乌苏里蝮）的系统发生关系，研究中使用了３３个蝮蛇标本和２个草原蝰标本，用１１个引物对样品分别作ＲＡＰＤ扩增，获得７２个扩增片段。     

RAPD在双歧杆菌菌种鉴定及分型研究中的应用

采用近年来兴起的一种新的分子生物学分型技术ＲＡＰＤ即ＡＰ－ＰＣＲ对２０株４种不不双歧杆菌进行了基因指纹图谱的构建,其结果不同双歧杆菌种间存在同源性和多态性。本研究结果表明ＲＡＰＤ技术可用于双歧杆菌菌种鉴定及分型。     

随机扩增多态性DNA技术在林业中的应用

１ＲＡＰＤ的原理随机扩增多态性ＤＮＡ（ＲａｎｄｏｍＡｍｐｌｉｆｉｅｄＰｏｌｙｍｏｒｐｈｉｃＤＮＡ,ＲＡＰＤ）系１９９０年由美国科学家采用ＰＣＲ技术发展起来的。在发现ＲＡＰＤ多态性的同时,也证明了ＲＡＰＤ标记的分离符合孟德尔独立分配规律,从而确立了ＲＡ...     

苍术DNA分离及RAPD遗传多样性分析

用新鲜的或－７５℃保存的苍术〔Ａｔｒａｃｔｙｌｏｄｅｓｌａｎｃｅａ（Ｔｈｕｎｂ．）ＤＣ．〕叶片提取ＤＮＡ,产量分别为４０ｎｇ／ｍｇ鲜重和１０ｎｇ／ｍｇ鲜重左右,分离出的ＤＮＡ的ＯＤ２６０／ＯＤ２８０值均大于１９,可用于ＲＡＰＤ试验。用８种引物对苍术４个居群的ＤＮＡ进行扩增,单个１０碱基的引物扩增出的ＲＡＰＤ标记在１～１５之间,多态位点百分率南苍术为４７５０％,北苍术为４５４０％,遗传相似度值南苍术为０８５,北苍术为０８７。结果表明,南苍术的遗传多样性略高于北苍术。     

人体蠕形螨的DNA提取与随机引物PCR检测

【目的】探索人体毛囊蠕形螨和皮脂蠕形螨DNA的提取方法。【方法】采用液氮反复冻融研磨法破碎螨体细胞, 选用改良小昆虫DNA提取法、碱裂解法和试剂盒提取法, 分别提取冻存时间在5个月内和8~10个月的毛囊蠕形螨和皮脂蠕形螨基因组DNA, 并用随机引物PCR方法进行检测。【结果】蛋白核酸测定仪检测结果显示, 试剂盒法提取的DNA纯度较高、量较多, 明显优于改良小昆虫法和碱裂解法。随机引物扩增结果显示清晰的DNA指纹图谱, 两种人体蠕形螨DNA指纹具有明显差异。蠕形螨冻存时间影响DNA提取的量, 但对DNA提取的纯度和RAPD指纹图谱影响较小。不同DNA提取方法提取的同一种蠕形螨DNA指纹图谱基本相似, 试剂盒法和改良小昆虫法提取的DNA样本条带多而清晰, 碱裂解法提取的样本条带少而模糊。【结论】液氮反复冻融研磨法破碎蠕形螨细胞是有效的, 蠕形螨冻存时间不宜超过6个月, 试剂盒提取法是提取蠕形螨DNA的好方法。RAPD技术可以用于这两种人体蠕形螨DNA分子水平上的检测和分类。     

Effect of DNA preparation from laver (Porphyra yezoensis) thalli on reproducibility of RAPD (random amplified polymorphic DNA) patterns

Laver (Porphyra yezoensis) DNAs were extracted from thalli with five different procedures and used for RAPD (random amplified polymorphic DNA) analysis as templates. Restriction enzyme-digestive DNAs were obtained with all procedures examined. However, RAPD patterns generated with these DNAs appeared highly irreproducible and were considerably different from each other. When DNAs purified with CsCl gradient centrifugation were used for RAPD analysis as templates, highly reproducible RAPD patterns were obtained, suggesting that unpurified DNAs extracted from thalli with all five extraction procedures contained an excess of RNA, polysaccharides and/or other materials which affected the RAPD reproducibility. Thus, results indicated that purification of DNA is essential to produce reproducible RAPD patterns of Porphyra DNA. This revised version was published online in June 2006 with corrections to the Cover Date.     

Detection of Pseudomonas putida B in the rhizosphere by RAPD

A method was developed for the detection of Pseudomonas putida B MM12 released into the rhizosphere of non-sterile barley, using a Random Amplified Polymorphic DNA (RAPD)-generated probe for hybridization with RAPD products generated from DNA extracted from the rhizosphere. The detection procedure involves extraction of rhizosphere bacteria by sonication, extraction of DNA by boiling, RAPD and Southern hybridization with RAPD products and the selected probe. The level of detection of MM12 was at least 1·9×10⁴ cells g⁻¹ barley root. MM12 was detected in rhizosphere when it constituted as little as 0·5% of the culturable population.     

Ready-to-use DNA extracted with a CTAB method adapted for herbarium specimens and mucilaginous plant tissue

This report summarizes major changes in previously published protocols for DNA extraction to improve the quality of DNA extracted from plants. Here, we highlight the critical modifications in the original protocols. The efficiency of these changes results in high-quality DNA ready to use in a variety of phytogenetically distant plant families, in particular species with mucopolysaccharides. The DNA obtained can be used without further purification in various molecular biology assays, including direct sequencing and AFLP and RAPD (random-amplified polymorphic DNA) analyses. The effectiveness of this method is proven by the amplification and sequencing of PCR products of up to 1 kb with DNA extracted from herbarium tissue ≥60 years old. This versatility is not usually found in DNA extraction protocols. In addition, this method is quick, adaptable to standard laboratories, and most important, safer and more cost-effective.     

红松RAPD实验中各组成成份含量对实验结果的影响

本文采用Ｐｒｏｍｅｇａ的ＰＣＲ反应系统,以从红松幼叶中提取的总ＤＮＡ为模板,进行随机扩增多态ＤＮＡ（ＲＡＰＤ）实验。     

Rapid DNA extraction methods and new primers for randomly amplified polymorphic DNA analysis of Giardia duodenalis.

A randomly amplified polymorphic DNA (RAPD) procedure using simple genomic DNA preparation methods and newly designed primers was optimized for analyzing Giardia duodenalis strains. Genomic DNA was extracted from in vitro cultivated trophozoites by five freezing-thawing cycles or by sonic treatment. Compared to a conventional method involving proteinase K digestion and phenol extraction, both freezing-thawing and sonication were equally efficient, yet with the advantage of being much less time- and labor-intensive. Five of the 10 tested RAPD primers produced reproducible polymorphisms among five human origin G. duodenalis strains, and grouping of these strains based on RAPD profiles was in agreement among these primers. The consistent classification of two standard laboratory reference strains, Portland-1 and WB, in the same group confirmed previous results using other fingerprinting methods, indicating that the reported simple DNA extraction methods and the selected primers are useful in RAPD for molecular characterization of G. duodenalis strains.     

一种提取麦红吸浆虫基因组DNA的方法

介绍一种可从单头麦红吸浆虫Sitodiplosis mosellana(Gehin)成虫、幼虫和蛹成功地提取基因组DNA的方法。经检测提取DNA样品浓度为100～200ng/μL,纯度在1.4～1.6范围之间,以此为模板,可顺利地进行RAPD引物扩增。该提取方法简单、安全、经济,可为小型昆虫基因组DNA的提取提供参考。     

栗属植物基因组DNA的提取及RAPD、SSR分析

以中国板栗、茅栗和锥栗的叶片为材料,提取栗属的DNA。针对栗属植物富含多酚类等次生物质的特点对栗属DNA的提取方法进行了改良,采用CTAB-蛋白酶K法加重复抽提,去除栗属植物中的相关次生物质,获得了高质量的DNA。所提取的DNA完全满足PCR、RAPD、SSR等一些分子生物学实验要求。     

大血藤DNA提取及RAPD研究初探

以浙江天台山大血藤为材料,对其DNA提取和RAPD分子标记方法进行了研究。结果表明:所采用的经改进的SDS提取法可获得高质量的大血藤DNA,分子量大于23kb,可满足RPAD扩增;用15个不同的随机引物对所提取的12个个体的大血藤DNA进行了RAPD分子标记分析,其中14个引物扩增产物具有多态性。建立了大血藤DNA提取和RAPD标记的分析程序,为RAPD分析应用于大血藤遗传多样性研究打下了良好的基础。     

丝状真菌组织DNA的提取

用CTAB法直接从丝状真菌的新鲜菌丝中提取DNA,并将所提取DNA进行随机引物多态性扩增(RAPD),得到了较清晰的扩增图谱,证明该法为提取真菌DNA以用于分子生物学研究的一种有效方法。