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以大肠杆菌(E.coli)为对象,采用环介导恒温扩增技术(LAMP)对其扩增,在实验室自制的毛细管电泳-诱导荧光平台上建立了LAMP产物的检测新方法。引物F3,B3,FIP,BIP扩增的E.coli LAMP产物大小为240 bp。优化的毛细管电泳条件为:毛细管有效长度/总长度(10 cm/15 cm),筛分介质溶液为0.5%羟乙基纤维素(1 300 K),电场强度(100 V/cm),进样条件(100 V/cm,1.0 s)。毛细管电泳时,DNA长度在100~500 bp范围内与其迁移时间呈线性关系,相关系数为0.996。在相同毛细管电泳条件下对E.coli LAMP产物进行分析,并利用这种线性关系在电泳图中对E.coli LAMP产物与假阳性产物做区分,结果表明,毛细管电泳技术不仅可在15 min内实现LAMP产物及附加产物的快速检测,而且可快速区分LAMP阳性及假阳性实验产物。采用建立的毛细管电泳快速检测LAMP产物的方法,对AB0174 E.coli基因实施了LAMP,结果表明该方法适合DNA LAMP产物的快速检测。 相似文献
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建立了毛细管电泳快速检测牙龈卟啉单胞菌(P.g)、齿垢密螺旋体(T.d)、福赛斯坦纳菌(T.f)3种牙周病病原菌种的方法。紫外分光光度法表明,采用无菌吸潮纸尖及PBS缓冲液可以实现牙周病原菌的快速有效提取。对提取的牙周病病原菌种做多聚酶链式反应(PCR)与多重PCR反应,最后以20 cm长,75μm内径的石英毛细管作为分离通道,分离电压4000 V,1.2%羟乙基纤维素(HEC,250 K)为筛分介质,牙周病病原菌菌种P.g,T.d,T.f PCR及多重PCR产物12 min内得到较好分离。结果表明,毛细管电泳与PCR技术相结合,可以实现牙周病原菌种快速鉴定,且检测限低至4.80×10-11ng/μL。方法已用于牙周病原菌菌种的快速检测。 相似文献
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The doctor-blade method is investigated for the preparation of Cu2ZnSnS4 films for low-cost solar cell application.Cu2ZnSnS4 precursor powder,the main raw material for the doctor-blade paste,is synthesized by a simple ball-milling process.The doctor-bladed Cu2ZnSnS4 films are annealed in N2 ambient under various conditions and characterized by X-ray diffraction,ultraviolent/vis spectrophotometry,scanning electron microscopy,and current-voltage(J-V) meansurement.Our experimental results indicate that(i) the X-ray diffraction peaks of the Cu2ZnSnS4 precursor powder each show a red shift of about 0.4°;(ii) the high-temperature annealing process can effectively improve the crystallinity of the doctor-bladed Cu 2 ZnSnS 4,whereas an overlong annealing introduces defects;(iii) the band gap value of the doctor-bladed Cu 2 ZnSnS 4 is around 1.41 eV;(iv) the short-circuit current density,the open-circuit voltage,the fill factor,and the efficiency of the best Cu2ZnSnS4 solar cell obtained with the superstrate structure of fluorine-doped tin oxide glass/TiO2/In2S3/Cu2ZnSnS4/Mo are 7.82 mA/cm2,240 mV,0.29,and 0.55%,respectively. 相似文献
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Geometric, electronic and vibrational properties of the most stable and energetically favourable configurations of indium oxide clusters InmOn (1 ≤ m, n ≤ 4) are investigated using density functional theory. The lowest energy geometries prefer the planar arrangement of the constituent atoms with a trend to maximize the number of ionic In-O bonds. Due to the charge transfer from In to O atoms, the electrostatic repulsion occurs between the atoms with the same kind of charge. The minimization of electrostatic repulsion and the maximization of In-O bond number compete between each other and determine the location of the isometric total energy. The most stable linear In-O-In-O structure of In2O2 cluster is attributed to the reduced electrostatic repulsive energy at the expense of In-O bond number, while the lowest energy rhombus-like structure of In2O3 cluster reflects the maximized number of In-O bonds. Furthermore, the vibrational frequencies of the lowest energy clusters are calculated and compared with the available experimental results. The energy gap and the charge density distribution for clusters with varying oxygen/indium ratio are also discussed. 相似文献
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以羟乙基纤维素为筛分介质,在直流、方波脉冲、反向脉冲电场中对0.1~10.0 kbp范围的DNA样品进行分离,改变脉冲电场调制深度,探讨电场方式对毛细管电泳分离DNA的影响.研究发现,其它实验条件一定时:(1)直流电场下,小片段DNA(<1.0 kbp)可以被有效分离,大片段DNA(>1.0 kbp)迁移时几乎重叠在一起;(2)方波脉冲电场下,增大调制深度可提高大片段DNA(>1.0 kbp)分离效果,但降低了部分小片段DNA(0.6~1.0 kbp)的分离度;(3)反向脉冲电场下,可以实现0.1~8.0 kbp范围内各个DNA片段的有效分离,改变调制深度会影响样本DNA的分离时间.并将反向脉冲电场应用于毛细管电泳分离λ-DNA的EcoT14 I/Bgl II限制性内切酶酶切片段.结果表明,反向脉冲毛细管电泳技术具有快速、准确、重复性高等特点,可用于宽分子量范围DNA片段分离. 相似文献
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The doctor-blade method is investigated for the preparation of Cu2ZnSnS4 films for low-cost solar cell application. Cu2ZnSnS4 precursor powder, the main raw material for the doctor-blade paste, is synthesized by a simple ball-milling process. The doctor-bladed Cu2ZnSnS4 films are annealed in N2 ambient under various conditions and characterized by X-ray diffraction, ultraviolent/vis spectrophotometry, scanning electron microscopy, and current-voltage (J-V) meansurement. Our experimental results indicate that (i) the X-ray diffraction peaks of the Cu2ZnSnS4 precursor powder each show a red shift of about 0.4°; (ii) the high-temperature annealing process can effectively improve the crystallinity of the doctor-bladed Cu2ZnSnS4, whereas an overlong annealing introduces defects; (iii) the band gap value of the doctor-bladed Cu2ZnSnS4 is around 1.41 eV; (iv) the short-circuit current density, the open-circuit voltage, the fill factor, and the efficiency of the best Cu2ZnSnS4 solar cell obtained with the superstrate structure of fluorine-doped tin oxide glass/TiO2/In2S3/Cu2ZnSnS4/Mo are 7.82 mA/cm2, 240 mV, 0.29, and 0.55%, respectively. 相似文献
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本实验以羟乙基纤维素(HEC)为筛分介质,以100~1500 bp DNA ladder为分离对象,系统地研究了直流电场下毛细管电泳时DNA分离特性.论文考察了DNA迁移淌度及分离度随HEC溶液浓度和分子量、毛细管两端电场强度(E)、毛细管有效长度(le)及其内径形状、背景电解液(BGE)温度等因素变化规律.研究发现:(1)当筛分介质HEC浓度高于其阈值浓度c*时,HEC分子量越大,相邻DNA片段之间淌度差越大,HEC浓度越高,其迁移淌度越低;(2)对于相邻的DNA片段,le在一定范围内,其分离度随le增大而线性升高;(3)毛细管有效长度一定时,DNA淌度随毛细管侧面积与截面积之比R增大而升高,分离效率提高;(4)BGE温度升高,DNA在筛分介质中扩散效应增强,迁移淌度变大,相邻DNA片段间分离度减小.根据以上结论,在直流电场下毛细管电泳φ×174-Hirc II限制性酶切片段,并实现了其高分离度、快速分离. 相似文献
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为克服传统平板凝胶电泳法操作步骤繁琐,实验时间长,以及采用紫外光源可能对人体造成伤害等缺陷,该文研制了一种基于电泳芯片的快速凝胶电泳仪,仅需DNA点样于电泳芯片后,将电泳芯片置于该电泳仪便可实现DNA样品快速分离及实时在线成像检测。以20、50、100 bp DNA ladder为检测对象,在仪器内对其分离效果进行验证及优化,其最佳电泳条件为电压100 V/cm,琼脂糖质量分数分别为2. 5%、1. 0%、1. 4%。结果表明在14 min内可以实现3种DNA样品的高效分离,DNA迁移距离与分子量的相关系数均高于0. 9,且该仪器具有较高的稳定性。 相似文献