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1.
高粱胞质雄性不育及保持系的RAPD技术优化   总被引:2,自引:0,他引:2  
以高梁细胞质雄性不育系和保持系A1T623A/B,A2V2A/B为材料,考察了Mg^2 ,Taq酶,dNTP,引物,模板等的用量、循环参数及不同的PCR仪对随机扩增多态性DNA(RAPD)反应的影响。结果发现:在25μL体系中,MgCl22.0mmol/L,Taq酶0.75U(1U=1μmol/min),dNTP0.15mmol/L,引物16.5ng,模板30ng,明0.001%,KCl50mmol。L,Tris10mmol/L(pH8.3)为最佳反应混合物组合。对PeltierThermalCycler200而言,94℃预变性5min,94℃预变性5in,94℃变性1min,35℃退火1min,72℃延伸2min,最后72℃保温5min,共计40个循环为最佳扩增条件;对PerkinElmerCetusDNAtThermalCycler-480而言,采用94℃预变性3min后,前3个循环,94℃变性1min,35℃退火1min,72℃延伸2min,后37个循环,94℃变性30s,36℃退火45s,72℃延伸45s,72℃延伸1min,最后72℃保温5min的循环程序可获得理想扩增。并阐述了优化RAPD实验体系的设计原则。  相似文献   

2.
以干用辣椒幼苗为材料,研究了干用辣椒RAPD分析过程中的Taq酶、Mg^2+,dNTPs、引物、模板DNA浓度、退火温度等影响因素,建立适合干用辣椒RAPD反应的PCR体系,即25μL反应体系中含有解体Taq酶1.5U、Mg^2+2.5mmol/L,dNTPs0.6mmol/L、引物0.8μmol/L、模板DNA60ng.扩增程序为:94℃预变性4min;94℃变性1min,37℃退火1min,72%延伸1.5min,40个循环;最后72℃副延伸5min.  相似文献   

3.
通过对水稻RAPD反应条件比较研究,建立了对除草剂敏感致死水稻RAPD的最适反应体系和反应扩增程序,即在25μl反应体系中,含10 mM Tris-HCl(pH8.0),10 mM KCl,8 mM(NH4)2SO4,0.05%NP-40,2.0 mM MgCl2,0.1 mM(each)dTNPs,20 ng引物,20ng基因组DNA,1.5 U的Taq酶.反应扩增程序为:94℃变性2 min;扩增40个循环,每循环包括:94℃变性10 s,40℃退火30 s,72℃延伸1.5 min;最终延伸5 min.  相似文献   

4.
菜心基因组DNA提取及RAPD反应体系的优化   总被引:10,自引:0,他引:10  
用改良的CTAB法提取菜心基因组DNA,并利用紫外分光光度法测定其纯度和含量,用琼脂糖凝胶电泳法检测DNA的降解状况和纯度.结果表明:此法提取的DNA具有典型的天然DNA分子的标准紫外吸收光谱特点, 且适于进行RAPD分析.RAPD分析的优化反应体系为:25μL的反应液中含有0.8 U的Taq酶,0.28 μmol/L primer,30 ng的DNA,2.0 mmol/L Mg2 ,0.20 mmol/L dNTPs.PCR扩增循环为94℃、5 min;94℃、1 min; 36℃、1 min;72℃、2 min,37个循环;72℃延伸10 min.  相似文献   

5.
欧洲甜樱桃RAPD反应体系的建立与优化   总被引:1,自引:0,他引:1  
以欧洲甜樱桃萨米托品种为研究对象,利用改进的CTAB法提取基因组DNA,并通过单因素多水平梯度试验,筛选了模板、Mg2+、TaqE、dNTPs和随机引物的浓度及用量,建立了欧洲甜樱桃RAPD技术扩增体系.研究结果表明,在25出反应体系中,各组分最佳的浓度分别为:10×Buffer2.5μl,2.5mmol·L^-1Mg2+2.5μl,Taq E0.06U·μl^-1,0.2mmol·L^-1dNTPs,0.2μmol·L^-1Primer,模板DNAl.2ng·μl^-1.PCR循环程序为:94℃预变性4min。94℃变性40s,36℃退火45s,72℃延伸1min,共36个循环,最后72℃延伸7min.  相似文献   

6.
刘金文  沙伟  滕兆岩 《广西科学》2004,11(2):148-150,160
采用正交设计和响应曲面设计(RSD)优化适合于芦苇[Phragmites australis (Cav.)Trin.ex steud]增多态DNA(RAPD)的反应体系,结果确定芦苇RAPD反应体系的最佳方案为:在PCR扩增程序,冷启动,94℃变性1min,36℃复性90s,72℃延伸2min,40个循环;72℃延伸7min,4℃保存;25μl反应体系包括模板DNA 60ng、三磷酸脱氧核苷酸(dNTP)7.5mmol、Taq酶1.5u、引物5pmol。  相似文献   

7.
流苏石斛ISSR-PCR反应体系的优化   总被引:2,自引:0,他引:2  
以流苏石斛为材料,对影响ISSR-PCR扩增结果的因素进行了探讨,建立了适合流苏石斛ISSR-PCR分析的最佳反应体系.在20μL反应体系中含1×buffer,2.0 mmol/L Mg2+,1.0 mmol/L dNTP s,1.0μmol/L引物,0.5 UTaqDNA聚合酶,10 ng模板DNA.扩增程序为:94℃预变性5 min;94℃变性45 s,50℃退火45 s,72℃延伸45 s,共35个循环;最后72℃延伸7 min.  相似文献   

8.
为获得铁皮石斛RAPD的最佳反应体系,本文对影响RAPD反应的模板含量、Mg2+浓度、dNTP浓度、Taq酶浓度、扩增程序及退火温度等多种因子进行了构建与优化.通过各因子的组合研究,我们得到铁皮石斛RAPD反应最适扩增体系是:25μLPCR反应体积,10×Buffer 2.5μL,2.5 mmol/L MgC l2,dNTP 2.5μL(各2.5mmol/L),模板DNA为30 ng,Taq聚合酶1.5 U,随机引物12 pmol,ddH2O 14.2μL.最佳扩增程序:94℃预变性3m in,94℃变性50 s,38℃复性1 m in,72℃延伸2 m in,循环40次;最后72℃延伸5 m in.4℃保存.  相似文献   

9.
山药基因组DNA的提取和RAPD反应条件的优化   总被引:5,自引:0,他引:5  
研究了山药基因组DNA的提取方法;对Mg2+浓度、dNTPs浓度、引物浓度和Taq酶用量进行了优化,建立了最佳的RAPD反应体系:在25μL反应体系中Mg2+浓度为2.0 mmol.L-1,dNTPs浓度为0.25 mmol.L-1,引物浓度为20 pmol.L-1,Taq酶1.5 U,模板DNA为200 ng.扩增程序为:95℃预变性7 min;94℃变性30 s,36℃退火45 s,72℃延伸2 min,共35个循环;72℃终延伸10 min.  相似文献   

10.
以狗脊蕨新鲜叶片为材料,使用改良的CTAB法提取其基因组DNA,通过单因子实验优化了各反应物的浓度,即在25μL总反应体系,Mg2 (2.5 mmol/L),dNTPs(200 mmol/L),DNA(15 ng),Taq酶(1.5 U),引物(0.15μmmol/L),扩增程序为:94℃预变性1 min,然后94℃变性1 min,39℃退火1min,72℃延伸2 min,40个循环,最后72℃延伸10 min,4℃保存.狗脊RAPD条件优化为分析狗脊蕨的遗传多样性打下稳定的基础.  相似文献   

11.
Language markedness is a common phenomenon in languages, and is reflected from hearing, vision and sense, i.e. the variation in the three aspects such as phonology, morphology and semantics. This paper focuses on the interpretation of markedness in language use following the three perspectives, i.e. pragmatic interpretation, psychological interpretation and cognitive interpretation, with an aim to define the function of markedness.  相似文献   

12.
The discovery of the prolific Ordovician Red River reservoirs in 1995 in southeastern Saskatchewan was the catalyst for extensive exploration activity which resulted in the discovery of more than 15 new Red River pools. The best yields of Red River production to date have been from dolomite reservoirs. Understanding the processes of dolomitization is, therefore, crucial for the prediction of the connectivity, spatial distribution and heterogeneity of dolomite reservoirs.The Red River reservoirs in the Midale area consist of 3~4 thin dolomitized zones, with a total thickness of about 20 m, which occur at the top of the Yeoman Formation. Two types of replacement dolomite were recognized in the Red River reservoir: dolomitized burrow infills and dolomitized host matrix. The spatial distribution of dolomite suggests that burrowing organisms played an important role in facilitating the fluid flow in the backfilled sediments. This resulted in penecontemporaneous dolomitization of burrow infills by normal seawater. The dolomite in the host matrix is interpreted as having occurred at shallow burial by evaporitic seawater during precipitation of Lake Almar anhydrite that immediately overlies the Yeoman Formation. However, the low δ18O values of dolomited burrow infills (-5.9‰~ -7.8‰, PDB) and matrix dolomites (-6.6‰~ -8.1‰, avg. -7.4‰ PDB) compared to the estimated values for the late Ordovician marine dolomite could be attributed to modification and alteration of dolomite at higher temperatures during deeper burial, which could also be responsible for its 87Sr/86Sr ratios (0.7084~0.7088) that are higher than suggested for the late Ordovician seawaters (0.7078~0.7080). The trace amounts of saddle dolomite cement in the Red River carbonates are probably related to "cannibalization" of earlier replacement dolomite during the chemical compaction.  相似文献   

13.
何延凌 《科技信息》2008,(4):258-258
Language is a means of verbal communication. People use language to communicate with each other. In the society, no two speakers are exactly alike in the way of speaking. Some differences are due to age, gender, statue and personality. Above all, gender is one of the obvious reasons. The writer of this paper tries to describe the features of women's language from these perspectives: pronunciation, intonation, diction, subjects, grammar and discourse. From the discussion of the features of women's language, more attention should be paid to language use in social context. What's more, the linguistic phenomena in a speaking community can be understood more thoroughly.  相似文献   

14.
AcomputergeneratorforrandomlylayeredstructuresYUJia shun1,2,HEZhen hua2(1.TheInstituteofGeologicalandNuclearSciences,NewZealand;2.StateKeyLaboratoryofOilandGasReservoirGeologyandExploitation,ChengduUniversityofTechnology,China)Abstract:Analgorithmisintrod…  相似文献   

15.
理论推导与室内实验相结合,建立了低渗透非均质砂岩油藏启动压力梯度确定方法。首先借助油藏流场与电场相似的原理,推导了非均质砂岩油藏启动压力梯度计算公式。其次基于稳定流实验方法,建立了非均质砂岩油藏启动压力梯度测试方法。结果表明:低渗透非均质砂岩油藏的启动压力梯度确定遵循两个等效原则。平面非均质油藏的启动压力梯度等于各级渗透率段的启动压力梯度关于长度的加权平均;纵向非均质油藏的启动压力梯度等于各渗透率层的启动压力梯度关于渗透率与渗流面积乘积的加权平均。研究成果可用于有效指导低渗透非均质砂岩油藏的合理井距确定,促进该类油藏的高效开发。  相似文献   

16.
As an American modern novelist who were famous in the literary world, Hemingway was not a person who always followed the trend but a sharp observer. At the same time, he was a tragedy maestro, he paid great attention on existence, fate and end-result. The dramatis personae's tragedy of his works was an extreme limit by all means tragedy on the meaning of fearless challenge that failed. The beauty of tragedy was not produced on the destruction of life, but now this kind of value was in the impact activity. They performed for the reader about the tragedy on challenging for the limit and the death.  相似文献   

17.
There are numerous geometric objects stored in the spatial databases. An importance function in a spatial database is that users can browse the geometric objects as a map efficiently. Thus the spatial database should display the geometric objects users concern about swiftly onto the display window. This process includes two operations:retrieve data from database and then draw them onto screen. Accordingly, to improve the efficiency, we should try to reduce time of both retrieving object and displaying them. The former can be achieved with the aid of spatial index such as R-tree, the latter require to simplify the objects. Simplification means that objects are shown with sufficient but not with unnecessary detail which depend on the scale of browse. So the major problem is how to retrieve data at different detail level efficiently. This paper introduces the implementation of a multi-scale index in the spatial database SISP (Spatial Information Shared Platform) which is generalized from R-tree. The difference between the generalization and the R-tree lies on two facets: One is that every node and geometric object in the generalization is assigned with a importance value which denote the importance of them, and every vertex in the objects are assigned with a importance value,too. The importance value can be use to decide which data should be retrieve from disk in a query. The other difference is that geometric objects in the generalization are divided into one or more sub-blocks, and vertexes are total ordered by their importance value. With the help of the generalized R-tree, one can easily retrieve data at different detail levels.Some experiments are performed on real-life data to evaluate the performance of solutions that separately use normal spatial index and multi-scale spatial index. The results show that the solution using multi-scale index in SISP is satisfying.  相似文献   

18.
本文叙述了对海南岛及其毗邻大陆边缘白垩纪到第四纪地层岩石进行古地磁研究的全部工作过程。通过分析岩石中剩余磁矢量的磁偏角及磁倾角的变化,提出海南岛白垩纪以来经历的构造演化模式如下:早期伴随顺时针旋转而向南迁移,后期伴随逆时针转动并向北运移。联系该地区及邻区的地质、地球物理资料,对海南岛上述的构造地体运动提出以下认识:北部湾内早期有一拉张作用,主要是该作用使湾内地壳显著伸长减薄,形成北部湾盆地。从而导致了海南岛的早期构造运动,而海南岛后期的构造运动则主要是受南海海底扩张的影响。海南地体运动规律的阐明对于了解北部湾油气盆地的形成演化有重要的理论和实际意义。  相似文献   

19.
In the 19th century the society was controlled by men, and women were just appendants of them, they had not any rights and freedom. But Jane was an exception, she showed some characteristics of early feminist. Jane showed her characteristics of feminism in three aspects: rebellion, equality, and independence. These characteristics were helpful to her success, and feminism is the only way out for women of that time.  相似文献   

20.
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