ARMS实时PCR基因分型特异性的研究

以原癌基因K—ras基因12位密码子的4种基因型(野生型为GGT，突变型为AGT、TGT、CGT)质粒DNA为研究模型．采用SYBR Green I为实时PCR指示剂，进行ARMS实时PCR基因分型特异性的研究．结果表明：在和模板匹配的ARMS引物3′端附近引人故意错配碱基．将ARMS引物浓度稀释10倍，并对传统三步PCR循环进行改良．即根据目的产物和引物二聚体的熔点差异．在传统延伸步骤之后增加一步恒温80℃检测荧光步骤。可消除引物二聚体的影响．使ARMS引物的特异性增强．得到清晰的基因分型结果．实验表明这种基于SYBR Green I的ARMS实时PCR基因分型方法，无需合成探针，实验设计简单．是一种快速、简便、经济、特异性好的基因分型方法，可推广用于其他基因的分型。并可望成为应用于临床的基因诊断技术之一．     

实时荧光定量PCR的原理及应用研究进展

实时荧光定量PCR（real-time fluorescent quantitative polymerase chain reaction,FQ-PCR）是在定性PCR技术基础上发展起来的核酸定量技术,已成为分子生物学和其它领域的重要技术工具。本文对实时荧光定量PCR技术的原理及其在食品、医学、环保领域的应用进行综述。     

DNA检测方法在鱼类物种鉴定中的应用

总结了近年来在鱼类物种鉴定中常用的DNA序列分析、DNA指纹技术、物种特异性聚合酶链式反应和实时荧光PCR技术等DNA检测方法,同时对各类方法的特点和局限性进行了分析．     

高粱BTx623幼苗多酚氧化酶Real-time PCR引物扩增效率的检测

介绍了一种检测实时荧光定量PCR技术引物扩增效率的方法,以期为进一步应用实时荧光定量PCR奠定基础。同时通过实时荧光定量PCR技术,研究高粱BTx623幼苗中的多酚氧化酶基因SbPPO的表达情况,结果发现SbPPO2,SbPPO5,SbPPO6的相对表达量较高,这3个SbPPO可能在高粱BTx623幼苗中起到重要功能。     

荧光定量PCR技术及其在预防兽医学研究中的应用

荧光定量PCR（fluorescencer quantitive polymerase chain reaction,FQ-PCR）技术是20世纪90年代中期发展起来的一种新型核酸定量技术。该技术具有实时监测、快速、灵敏、精确等特点,是对原有PCR技术的革新,扩大了PCR的应用范围。本文综述了FQ-PCR技术的基本原理及其在预防兽医学研究中的应用。     

快速敏捷检测人乳头瘤病毒基因型的实时荧光PCR方法

宫颈癌是女性最常见的一种恶性肿瘤,研究表明人乳头瘤病毒(HPV)感染与宫颈癌的发生密切相关.采用实时荧光PCR技术对引起宫颈癌的两种主要HPV亚型HPV16和18型阳性质粒进行检测,在实验室中成功构建了HPV16和18型的检测体系以及体外模拟的双重感染检测体系,并对三例临床确诊为宫颈癌的标本进行实时荧光PCR验证检测,确定三例标本均为高浓度HPV16型感染,极易引起宫颈癌变,与医院诊断结果相符合.本实验为实时荧光PCR技术检测HPV16和18型的临床检验在实验室中成功地进行了探索,为实时荧光PCR技术在宫颈癌大规模筛查上的应用提供了研究基础.     

荧光定量PCR技术及其在动物病毒病定量检测中的应用

荧光定量PCR技术是一种核酸定量技术,该技术在PCR反应体系中加入荧光基团,利用荧光信号累积实时监测整个PCR进程,最后通过标准曲线对未知模板进行定量分析的方法.该技术具有特异性强、灵敏度高、重复性好、定量准确、速度快、全封闭反应等特点,因此在动物病毒病的检测中,荧光定量PCR技术不但能定量检测病原体感染的强弱和在机体内的分布,还具有灵敏、快速、省力的特点.     

3种松材线虫分子检测技术的比较分析

分别用特异性引物法、ITS-PCR-RFLP法和实时荧光PCR法对松材线虫进行了检测,通过检测时间、检测精度和操作情况等方面的比较得出:特异性引物PCR法操作简便、成本低,是目前的常规检测技术;ITS-PCR-PFLP法可以鉴定到小种,相比特异性引物法更具说服力,但耗时较多,适用于线虫种群分化研究和种类鉴定,而不适合作为一种快速检测技术;实时荧光PCR法耗时短,灵敏度高,操作简便安全,只是仪器设备要求较高,适合松材线虫的口岸检验检疫工作.     

食品中单增李氏菌实时荧光PCR检测鉴定方法的建立

运用实时荧光PCR(Real-time PCR)技术建立了对食品中单增李氏菌进行检测的快速方法，设计的引物和探针的序列特异性强，省去凝胶电泳的繁琐，降低了污染的可能性，在对26种病原菌进行检测过程中，运用实时荧光PCR法和经典培养法进行比较，结果表明用实时荧光PCR法检测单核细胞增多李斯特氏菌，更快速、敏感、特异性更高。     

PCR技术研究新进展

介绍了实时定量RT-PCR的原理、方法和应用,包括SYBR Green I、AmpliSensor、Lightcycle技术和Complex探针技术;同时还对原位PCR技术的原理、方法和应用,简并PCR的原理、方法和应用作了综述。     

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.     

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.     

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.     

Characterization of Ordovician carbonate reservoirs, southeastern Saskatchewan, Canada

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.     

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.     

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.     

橡胶颗粒沥青混合料低温抗裂性试验研究

以AC-13级配为基础,将橡胶颗粒代替部分集料掺入混合料中,以低温弯曲试验为评价方法对不同橡胶颗粒掺量下沥青混合料的低温抗裂性进行研究,并引入应变能密度值对混合料的低温抗裂性进行综合评价.试验结果表明:橡胶颗粒沥青混合料试件的破坏微应变均超过2 300,满足冬寒区的技术指标;无论是否掺加橡胶颗粒,随着温度的下降,沥青混合料破坏时的最大弯拉强度增大,弯拉应变降低,劲度模量增大;弯曲应变能密度在胶粒掺量为1%左右时具有较大的弯曲应变能密度值,此时橡胶颗粒沥青混合料具有较好的低温抗裂性.     

A computer generator for randomly layered structures

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

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.