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.     

赤松ISSR-PCR反应体系的建立与优化

为了建立赤松ISSR-PCR反应体系,利用正交试验分别对影响赤松PCR反应的dNTP浓度、Mg~(2+)浓度、Taq酶浓度、引物浓度、模板浓度进行了优化,并通过梯度PCR确定引物的最佳退火温度和循环次数.最终确定赤松ISSR-PCR最佳反应体系及扩增条件为:25μL反应体系中含2.5μL 10×PCR Buffer、3 mmol·L~(-1)dNTP、1.5 mmol·L~(-1)Mg~(2+)、0.5 U Taq酶、0.3μmol·L~(-1)引物、9 ng模板;扩增程序为:94℃预变性5 min,然后进行94℃变性30 s,55℃退火45 s(退火温度随引物不同而变化),72℃延伸2 min,35个循环,最后72℃延伸7 min.这一ISSR-PCR反应体系,为今后利用ISSR技术对赤松进行物种分类鉴定奠定了基础.     

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。     

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.     

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.     

濒危植物长柄双花木ISSR-PCR反应体系的优化

采用改进的CTAB法提取长柄双花木(Disanthus cercidifoliusvar.longipes)基因组DNA,并以此DNA为模板,对ISSR-PCR的反应体系进行了优化,建立了适合长柄双花木ISSR-PCR的反应体系和程序,即20μL的反应体系中,含模板DNA20 ng,Mg2+2.0 mmol.L-1,引物0.25μmol.L-1,dNTPs 0.20 mmol.L-1,TaqDNA聚合酶1.0 U;扩增程序:94℃预变性5 min,94℃变性30 s,48~54℃复性45 s;72℃延伸90 s;共36个循环;循环结束后,72℃延伸7 min.本研究为利用这一分子标记对长柄双花木进行遗传多样性分析奠定了基础.     

油松遗传多样性研究的ISSR—PCR体系优化与初步应用

以油松基因组DNA为模板,对影响ISSR-PCR的主要因素进行研究,建立了适宜于油松的ISSR-PCR优化体系及扩增程序;并采用lSSR标记对山西灵空山和陕西洛南2个油松种群的遗传多样性进行了分析,同时将此结果与使用RAPD标记的研究结果进行了对比.结果表明,在20μL反应体系中,模板DNA、Mg2+、dNTPs、Taq DNA聚合酶和引物5种主要成分的最适浓度分别为40 ng、2.5 mmol·L-1、0.20 mmol·L-1、1 U和0.5 μmol·L-1.扩增程序为:94℃预变性5 min;94℃变性30s,最适温度(54～56℃)退火45s,72℃延伸2min.共计35个循环;循环结束后72℃延伸7min,4℃保存.山西灵空山和陕西洛南2个油松种群的Nei's遗传多样性分别为0.3520和0.3514,前者的遗传多样性显著高于后者(P<0.001).使用ISSR标记与RAPD标记所获结果基本一致,但是lSSR标记优于RAPD标记.     

正交设计优化太子参ISSR-PCR反应体系

利用正交设计实验对影响太子参ISSR-PCR扩增的重要因素(d NTPs、引物、Taq酶、模板DNA浓度)进行优化,以建立太子参ISSR-PCR的优化反应的最佳体系,25μL反应体系中包含d NTPs300μmol/L,引物500nmol/L,Taq酶1U及模板DNA 50ng.适宜的扩增程序为94℃预变性5min;94℃变性40s,46℃退火45s,72℃延伸2min,41次循环;最后于72℃延伸7min,4℃进行保存.该正交体系的建立,将为太子参种质资源鉴定及遗传多样性的研究提供技术支持.     

爱玉ISSR反应体系的建立与优化

通过单因子试验和正交设计,对影响爱玉ISSR-PCR扩增效果的因素,如模板DNA用量、Taq酶用量、dNTPs浓度、引物浓度、延伸时间和循环次数等指标进行优化.实验确立了可用于爱玉ISSR-PCR分析最适宜的反应体系：20 μL反应体积中含1 ng模板DNA、1.0 U Taq酶、0.4 μmol/L引物和0.18 mmol/L dNTPs;PCR扩增程序为94 ℃预变性4 min;94 ℃变性45 s,53 ℃退火45 s,72 ℃延伸2.5 min,共35个循环;72 ℃延伸7min,4 ℃保存.应用该体系对14份爱玉种质进行扩增,证实了该体系的适用性和稳定性.     

罗汉果ISSR-PCR反应体系的建立

以罗汉果 ( Siraitia grosvenorii) DNA为材料 ,分析了模板 DNA,Mg2 + ,d NTPs,引物的浓度 ,Taq DNA聚合酶的用量以及循环次数对 ISSR-PCR扩增结果的影响 ,确立了稳定的、可重复的罗汉果 ISSR最佳反应体系和 PCR扩增参数 :在 2 5μL的 PCR反应体系中 ,含 2 0～ 5 0 ng模板 DNA,1 U Taq酶 ,1× PCR缓冲液 ,2 .0 mmol/L Mg Cl2 ,4种 d NTPs各 2 0 0 μmol/L,0 .5 μmol/L引物 ;PCR扩增程序为 94°C预变性 3 min,接着进行 40个循环 :94°C变性 1 min,5 2°C退火 5 0 s,72°C延伸 2 min,循环结束后 72°C延伸 7min.     

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.     

Design and analysis of generic LBS application developing platform

Location based services is promising due to its novel working style and contents.A software platform is proposed to provide application programs of typical location based services and support new applications developing efficiently. The analysis shows that this scheme is easy implemented, low cost and adapt to all kinds of mobile nework system.     

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