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.     

华山松ISSR反应体系的优化

为获得条带清晰、稳定和多态性高的华山松ISSR扩增结果,对引物、Mg~(2+)浓度和退火温度等因素进行了优化.确定了ISSR分析的最佳PCR条件:15μL反应体系中,10×PCR buffer反应缓冲液1.5μL,模板DNA 22.5 ng,Mg~(2+)2.2 mmol/L,d NTPs 0.23 mmol/L,引物0.93μmol/L,Taq DNA聚合酶0.027 U/μL.PCR反应程序为:经94℃预变性4min后,经过94℃变性30 s,Tm-4.8℃退火30 s,72℃延伸108 s共33个循环;循环结束后72℃延伸10 min.从44条ISSR引物中筛选出12条扩增稳定、多态性丰富的ISSR引物.对华山松ISSR实验优化体系的建立,为今后利用ISSR分子标记技术开展华山松种间遗传变异分析和构建遗传图谱等研究奠定了基础.     

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．     

应用RAPD技术构建绞股蓝DNA指纹图谱

以不同类型绞股蓝(Gynostemma Pentaphyllum,(Thunb)Makino)DNA为模板,进行RAPD反应条件的优化及RAPD结果分析.最终筛选的RAPD反应条件为:20μL PCR反应体系中包括了10×buffer2.0μL,Mg2 2.0 mmol/L,dNTP 0.2 mmol/L,引物0.5 mmol/L,Taq酶1U,模板DNA 50 ng.反应程序为94℃3 min→(94℃30 s→38℃40 s→72℃1 min)45个循环→72℃10min→4℃保持,并从66条随机引物中筛选出5条带型丰富、适合于绞股蓝分析的随机引物.以这5条随机引物对不同类型绞股蓝进行RAPD扩增,识别其特征性多态位点,建立分子标记检索表.     

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.     

大白菜RAPD扩增体系的优化研究

以大白菜3411－7自交系为材料，使用PE公司生产的Thermal Cycler480型PCR仪，对影响大白菜RAPD扩增的因素进行了研究，确定了模板，Mg^2 ,dNTPs，引物和Taq DNA聚合酶的适宜浓度和最佳的循环次数。实验结果表明，在25μL反应体系中使用20－60mg的模板DNA，1.5-2.0mmol/L的Mg^2 ,0.15-0.25mmol/L dNTPs,0.8-1.0μmol/L随机引物，1.0-1.5U Taq DNA聚合酶，在94℃下预变性5min后执行94℃ 1min,37℃ 1min,72℃ 2min，35个循环，最后72℃再延伸5min的条件下，大白菜的RAPD扩增效果较好。     

RAPD法研究穗花杉遗传多样性的反应条件优化

以改进CTAB法抽提的中国特有濒危植物穗花杉嫩叶总DNA为模板,进行随机扩增多态性DNA(RAPD)最佳条件优化,结果表明:RAPD分析最佳反应体系为10×Buffer缓冲液2μL,模板70ng,Mg2+2.0mmol/L,dNTPs0.2mmol/L,引物0.60μmol/L,Taq酶1.5U,PCR反应体积为20μL.最佳扩增程序为94℃5min,1个循环;94℃1min、37℃1min、72℃2min,40个循环;72℃10min,1个循环.     

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。     

新疆杏品种自交不亲和S-RNase基因PCR扩增体系的优化

以3个新疆杏品种的叶片为试材,以其叶片基因组DNA为模板,分别对PCR扩增体系中的10×Buffer(含Mg2+)、dNTP、上下游引物、TaqDNA聚合酶和退火温度5个因素设计6个梯度,研究了新疆杏品种自交不亲和S-RNase基因PCR扩增条件,建立其优化的PCR扩增体系。研究结果表明:其扩增程序为94℃预变性5min,94℃变性45s,52℃退火45s,72℃延伸2min,35个循环后72℃延伸10min,25μL反应体系10×Buffer(含Mg2+)2.5μL,dNTP0.08mmol/L,上下引物0.16μmol/L,DNA80ng,DNA聚合酶1μL,此为最优的反应体系。     

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.     

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.     

An Analysis of Catherine＇s Character——Wuthering Heights

Wuthering Heights, Emily Bronte＇s only novel, was published in December of 1847 under the pseudonym Ellis Bell. The book did not gain immediate success, but it is now thought one of the finest novels in the English language. Catherine is the key character of this masterpiece, because everybody and everything center on her though she had a short life. We can understand this masterpiece better if we know Catherine well.     

Lower Paleozoic oil relationships within Williston Basin, Canada

The Williston Basin is a significant petroleum province, containing oil production zones that include the Middle Cambrian to Lower Ordovician, Upper Ordovician, Middle Devonian, Upper Devonian and Mississippian and within the Jurassic and Cretaceous. The oils of the Williston Basin exhibit a wide range of geochemical characteristics defined as "oil families", although the geochemical signature of the Cambrian Deadwood Formation and Lower Ordovician Winnipeg reservoired oils does not match any "oil family". Despite their close stratigraphic proximity, it is evident that the oils of the Lower Palaeozoic within the Williston Basin are distinct. This suggests the presence of a new "oil family" within the Williston Basin. Diagnostic geochemical signatures occur in the gasoline range chromatograms, within saturate fraction gas chromatograms and biomarker fingerprints. However, some of the established criteria and cross-plots that are currently used to segregate oils into distinct genetic families within the basin do not always meet with success, particularly when applied to the Lower Palaeozoic oils of the Deadwood and Winnipeg Formation.     

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