高粱胞质雄性不育及保持系的RAPD技术优化

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。     

干用辣椒RAPD—PCR反应体系正交优化的研究

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．     

水稻基因组DNA的RAPD扩增研究

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.     

菜心基因组DNA提取及RAPD反应体系的优化

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．     

欧洲甜樱桃RAPD反应体系的建立与优化

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．     

采用正交设计和响应曲面设计法优化芦苇随机扩增多态DNA体系

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。     

流苏石斛ISSR-PCR反应体系的优化

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.     

铁皮石斛野生居群研究(Ⅳ):RAPD反应体系的构建与优化

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.     

山药基因组DNA的提取和RAPD反应条件的优化

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.     

狗脊蕨(Woodwardia japonica(L.F.)Sm)基因组DNA提取与随机扩增多态性DNA(RAPD)的优化

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.     

Markedness and its Interpretation in Language Use

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.     

Features of Women's language

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.     

低渗透非均质砂岩油藏启动压力梯度研究

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。     

On Tragic Consciousness In The Works By Ernest Hemingway

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.     

Achieving Unusual States of Matter Under High Pressure

正The periodicity of the elements and the non-reactivity of the inner-shell electrons are two related principles of chemistry,rooted in the atomic shell structure.Within compounds,Group I elements,for example,invariably assume the+1 oxidation state,and their chemical properties differ completely from those of the p-block elements.These general rules govern our understanding of chemical structures and reactions.Using first principles calcula-     

Exchange-Correlation and Electronic Excitation Energies from Pairing Matrix Fluctuations

We have developed an adiabatic connection to formulate the ground-state exchange-correlation energy in terms of pairing matrix linear fluctuations.This formulation of the exchange-correlation energy opens a new channel for density functional approximations based on the many-body perturbation theory.We illustrate the potential of such approaches with an approximation based on the particle-particle Random Phase Approximation(pp-RPA).This re-     

Density Functional Theory for the Response of Periodic Systems to Electric Fields Based on the Vector Potential Approach

正The electronic and nuclear(structural/vibrational)response of 1D-3D nanoscale systems to electric fields gives rise to a host of optical,mechanical,spectral,etc.properties that are of high theoretical and applied interest.Due to the computational difficulty of treating such large systems it is convenient to model them as infinite and periodic(at least,in first approximation).The fundamental theoretical/computational problem in doing so is that     

On the Thermodynamic Limit for Responses to Static Electromagnetic Fields

For molecular systems,the quantum-mechanical treatment of their responses to static electromagnetic fields usually employs a scalar-potential treatment of the electric field and a vector-potential treatment of the magnetic field.Although the potential for each field separately is associated with the choice of an(unphysical)origin,the precise choice of the origin for the electrostatic field has little consequences for the results.This is different for the     

Call for Papers

<正>"The Journal of Shanghai Normal University:Mathematics"is published by Shanghai Normal University as regular issues of The Journal of Shanghai Normal University each year from 2014 in English.The editors-in-chief of the issues are professors Yuhao Cong and Maoan Han.The Journal of Shanghai Normal University was started in 1958 with     

Journal of Donghua University ( English Edition) Contents of Volume 31

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