共查询到20条相似文献，搜索用时 468 毫秒
1.
采用等高锁状均质电场(CHEF)凝胶电泳技术,对蛹虫草无性型-蛹草拟青霉(Paecilomyces militaris Liang)进行电泳并分析其核型.以温汉逊氏酵母(Hansenula wingei)及粟酒裂殖酵母(Schizosaccharomyces pombe)菌株的染色体DNA大小作为分子量标记,3株蛹草拟青霉基因组中各包含7条染色体DNA,其大小都在2 Mb至5.7 Mb之间.实验结果表明各菌株间核型不完全相同,存在一定的差异,呈现多态性. 相似文献
2.
Grain separation loss monitoring system in combine harvester 总被引：5，自引：0，他引：5
Zhan Zhao Yaoming LiJin Chen Jiaojiao Xu 《Computers and Electronics in Agriculture》2011,76(2):183-188
Based on laboratory experimental results obtained with an axial threshing test-rig with tangential feeding, cumulative distribution functions of separated grain in axial and radial directions of threshing rotor were built. Based on the analysis of the relationship between grain separation loss and grain separation flux in an area under the concave, an indirect grain separation loss monitoring method is presented in this paper.Piezo-electric polyvinylidene fluoride (PVDF) film was selected as sensitive material to design a grain flux sensor. While grain and material-other-than-grain (MOG) separated in the monitoring area impact on piezo-electric PVDF films, different electric charges are generated. After signal progressing with a charge amplifier, frequency discrimination and wave shaping, the number of grain can be counted by a microcontroller (MCU) and the grain separation loss of combine harvester can be measured in real-time.Field test results indicated that the measurement errors of grain separation loss recorded by the monitoring system relative to the loss checked manually were less than 12%. 相似文献
3.
使用常规酚/氯仿抽提法(CTAB法)、离心柱法和磁珠吸附法,分别从菖蒲芋螺毒腺、肌肉及肝胰脏中提取基因组DNA,利用普通琼脂糖凝胶电泳与脉冲场电泳(PFGE)对基因组DNA片段进行分离,并检测提取DNA的大小和分布范围,同时,对脉冲场电泳条件进行了优化。结果表明,3种方法均可提取基因组DNA,其中CTAB法和离心柱法提取的毒腺DNA纯度高,产率大,可用于后续实验,而磁珠法产率适中且纯度不高,但离心柱法的DNA片段较小,大多在9 kb以下,而CTAB法提取的DNA片段较大,获得的20 kb以上的大片段量较多,更适合于大片段基因组文库的构建。3种芋螺组织中,肝胰脏产率最高但片段弥散有降解,毒腺DNA片段较大且集中,产率也较高,而肌肉的DNA片段产率最低且片段较小。因此,用CTAB法提取菖蒲芋螺毒腺的DNA更适合构建大片段基因组DNA文库。筛选高质量的菖蒲芋螺的DNA,利用优化的脉冲场电泳条件进行分离回收,获得了不同大小片段的DNA,为后续菖蒲芋螺不同载体系统的基因组文库构建奠定了基础。 相似文献
4.
[目的]建立1种利用毛细管电泳快速、高效检测限制性内切酶酶切产物的方法。[方法]以甲基纤维素(MC)为筛分介质,用PBR322/BsuRⅠDNA Marker为试验对象,研究筛分介质浓度、pH值、毛细管柱温度和电场强度对毛细管电泳法分离小片段双链DNA的影响,寻求最佳电泳条件,并将此方法用于检测MspⅠ内切酶的酶切产物。[结果]MC浓度对双链小片段DNA的分离度有较大影响。随MC溶液浓度的增加,分离度呈先增后减的趋势。毛细管电泳的最佳条件为:MC浓度为2.0%,pH值为8.0,毛细管柱温度15℃,电场强度为275 V/cm。在此条件下,对MspⅠ内切酶的酶切产物进行检测,在20 min内检测到3个酶切片段。[结论]与平板凝胶电泳相比,运用毛细管电泳检测小片段限制性内切酶酶切产物更高效。 相似文献
5.
A nanofluidic channel device, consisting of many entropic traps, was designed and fabricated for the separation of long DNA molecules. The channel comprises narrow constrictions and wider regions that cause size-dependent trapping of DNA at the onset of a constriction. This process creates electrophoretic mobility differences, thus enabling efficient separation without the use of a gel matrix or pulsed electric fields. Samples of long DNA molecules (5000 to approximately 160,000 base pairs) were efficiently separated into bands in 15-millimeter-long channels. Multiple-channel devices operating in parallel were demonstrated. The efficiency, compactness, and ease of fabrication of the device suggest the possibility of more practical integrated DNA analysis systems. 相似文献
6.
Electrophoretic separations of large DNA molecules by periodic inversion of the electric field 总被引：144，自引：0，他引：144
In gel electrophoresis, nucleic acids and protein-detergent complexes larger than a threshold size all migrate at the same rate. For DNA molecules, this effect can be overcome by the simple procedure of periodically inverting the electric field. Tuning the frequency of the field inversions from 10 to 0.01 hertz, makes it possible to resolve selectively DNA's in the size range 15 to greater than 700 kilobase pairs. 相似文献
7.
A theoretical analysis of the reptational motion of DNA in a gel that includes the effects of molecular fluctuations has been used to explain the main features found in experiments involving periodic inversion of the electric field. The resonance-like decrease of the electrophoretic mobility as a function of pulse duration is related to transient "undershoots" in the orientation of the molecule, in agreement with recent experimental data. These features arise from a delicate interplay of internal and center of mass motion of the molecules under pulsed field conditions, and are important for the separation of DNA molecules in the size range 0.2 to 10 million base pairs. 相似文献
8.
为构建小白链霉菌武夷变种(Streptomyces albulus var. wuyiensis)的细菌人工染色体(bacterial artificial chromosomes,BAC)文库,提取了小白链霉菌武夷变种高分子量DNA(high molecular weight DNA,HMW-DNA)。用低熔点琼脂糖(low melting point agarose,LMP)包埋小白链霉菌武夷变种菌丝球,胶块经溶菌酶、蛋白酶K、两种限制性内切酶(BamHI、Hind III)处理后,使用箝位匀强电场(CHEF)凝胶电泳检测DNA片段大小。电泳结果显示提取HWM DNA完整,满足构建BAC文库的条件,确定了最适酶切条件为Hind III0.2 U酶切10min,为小白链霉菌武夷变种功能基因的研究奠定了基础。 相似文献
9.
A novel instrument for separating large DNA molecules with pulsed homogeneous electric fields 总被引：10，自引：0，他引：10
A new instrument has been developed for the electrophoretic separation of large DNA molecules that can independently regulate the voltage of each of 24 electrodes and allow the magnitude, orientation, homogeneity, and duration of the electric field to be precisely controlled. Each parameter can be varied at any time during the electrophoretic process. Thus distinct sets of conditions can be combined to optimize the separation of various fragment sizes in a single run. Independent control of electrode voltage allows all of the fields to be generated with electrodes arranged in a closed contour, independent of a particular geometry. This device increases both the resolution in any size range and the speed of separation, especially for DNA molecules larger than 3 megabases. 相似文献
10.
A protein mixture can be separated in a flow column by application of a transverse electrophoretic field to carry faster-migrating components of the mixture into a fixed gel bed at one side of the column, so that slower components move along in a buffer stream. The field is then reversed to returnthe faster-moving components to the buffer stream. The cycle is repeated many times to complete the separation. 相似文献
11.
A physical map of the Escherichia coli K12 genome 总被引：125，自引：0，他引：125
A physical map of a genome is the structure of its DNA. Construction of such a map is a first step in the complete characterization of that DNA. The restriction endonuclease Not I cuts the genome of Escherichia coli K12 into 22 DNA fragments ranging from 20 kilobases (20,000 base pairs) to 1000 kilobases. These can be separated by pulsed field gel electrophoresis. The order of the fragments in the genome was determined from available E. coli genetic information and analysis of partial digest patterns. The resulting ordered set of fragments is a macrorestriction map. This map facilitates genetic and molecular studies on E. coli, and its construction serves as a model for further endeavors on larger genomes. 相似文献
12.
[目的]筛选一种适合鸡血藤DNA的提取方法。[方法]以鸡血藤嫩叶为材料,分别采用液氮-CTAB法、石英砂-CTAB法以及石英砂-SDS法提取基因组DNA,利用琼脂糖凝胶电泳和紫外分光光度计检测提取的DNA样品,对提取方法进行比较筛选。[结果]电泳检测中,石英砂-CTAB法所提DNA条带亮度最高,质量较好。紫外检测中,石英砂-CTAB法提取的DNA A260/A280值大于1.8,比其他两种方法提取的DNA蛋白质污染小,其提取的DNA纯度和得率均最高,液氮-CTAB法提取DNA得率居中,石英砂-SDS法提取DNA得率较低。[结论]该研究结果表明石英砂比液氮更能简便快速地将叶片研碎,CTAB提取缓冲液比SDS提取缓冲液可更有效地去除杂质,使细胞裂解、DNA析出。因此,石英砂-CTAB法更适合鸡血藤DNA的提取。 相似文献
13.
小麦叶片蛋白质组的2D-LC分离及Nano LC-MS/MS分析 总被引：1，自引：0，他引：1
【目的】二维液相色谱（2D-LC）与双向电泳（2-DE）技术具有互补性,本文旨在探讨利用2D-LC和纳升级液相色谱串联质谱（Nano LC-MS/MS）技术分离鉴定小麦叶片蛋白质组的新方法。【方法】小麦叶片蛋白经提取、脱盐后,进行第一维阴离子交换分离;收集洗脱组分进行SDS-PAGE分析,并对非高丰度蛋白组分进行第二维反相液相色谱分离;随机选择含较低吸收峰的部分组分经胰蛋白酶水解后进行Nano LC-MS/MS 分析;将串联质谱数据通过MASCOT搜索NCBInr和EST数据库,并对二级质谱获得的蛋白序列进行MS BLAST分析。【结果】经第一维阴离子交换色谱分离,收集得到15个组分,其中第15组分包含小麦叶片丰度最高的蛋白——1,5-二磷酸核酮糖羧化酶/加氧酶（RuBisCO）;其余14个组分经反相液相色谱分离,共收集1 551个组分,获得1 867个色谱峰;随机对其中6个含较低吸收峰的收集组分酶解和Nano LC-MS/MS 检测,9种蛋白得到鉴定。【结论】结果初步表明,利用2D-LC和Nano LC-MS/MS技术可有效地分离和鉴定小麦叶片蛋白质组,为更深入全面地研究小麦叶片蛋白质组奠定基础。 相似文献
14.
两种检测扁桃基因组RAPD扩增产物方法的比较分析 总被引：1，自引：1，他引：0
通过2;(w/v)的琼脂糖凝胶和8;(w/v)的聚丙烯酰胺凝胶电泳对扁桃品种基因组DNA的RAPD检测.结果表明:8;(w/v)聚丙烯酰胺凝胶优于2;(w/v)琼脂糖凝胶,主要表现在,8;聚丙烯酰胺凝胶对长度相差100 bp以下的DNA分子的分离较2;的琼脂糖凝胶电泳效果好;且8;聚丙烯酰胺凝胶的分辨范围广,在线性DNA分子(0.1～2.0 kb)范围内也能得到很好的分离效果;用8;的聚丙烯酰胺凝胶分离扁桃品种基因组DNA的RAPD扩增结果,表现出在扁桃品种间DNA水平上差异大,运用此方法提高了扁桃品种的鉴别能力. 相似文献
15.
Sedimentation field flow fractionation (SFFF) is a method for purifying and providing mass or size distribution information on samples containing particulates or soluble macromolecules. Since SFFF separations are based on simple physical phenomena related to first principles, molecular weight (or particle sizes) can be determined without calibration standards. SFFF is a gentle technique suited for fractionating biomolecules. Studies with the fragile lambda DNA (molecular weight, 33 X 10(6] and smaller supercoiled plasmids have shown that these materials are not altered during SFFF separation; molecular weights and conformation remain unchanged, and biological activity is not reduced. Recoveries of nucleic acids approach 100 percent. Typically, components with about 20 percent difference in mass can be separated essentially to baseline if required. Fractionation time is usually independent of molecular weight, and separations often can be carried out within an hour. 相似文献
16.
为探讨柴胡单体皂苷的分离条件,采用超声波提取,正丁醇萃取,硅胶GH柱层析分离,层析检测方法。结果表明乙酸乙酯：95%乙醇（8：2）分离出三个单体化合物,说明硅胶柱层析可用于柴胡皂苷的制备性分离。 相似文献
17.
以豌豆(Pisum sativumL.)苗叶片为试材,提取其类囊体膜并利用不同增溶剂研究PSI蛋白复合物的分布以及不同凝胶浓度的DOC-PAGE分离特性。结果表明:6%的分离胶对PSI具有较好的分离特性;低浓度的毛地黄皂苷能够增溶间质类囊体上的PSI蛋白复合物,并能够获得大量的含有PSI的类囊体膜碎片。高浓度的毛地黄皂苷能够获得全部的PSI复合物;Tween-20能够分离不同区域的类囊体膜,结合毛地黄皂苷处理均能够获得PSI复合物。证明了PSI异质性的存在。 相似文献
18.
以豌豆（Pisum sativum L．）苗叶片为试材，提取其类囊体膜并利用不同增溶剂研究PSI蛋白复合物的分布以及不同凝胶浓度的DOC—PAGE分离特性。结果表明：6％的分离胶对PSI具有较好的分离特性；低浓度的毛地黄皂苷能够增溶间质类囊体上的PSI蛋白复合物，并能够获得大量的含有PSI的类囊体膜碎片。高浓度的毛地黄皂苷能够获得全部的PSI复合物；Tween-20能够分离不同区域的类囊体膜，结合毛地黄皂苷处理均能够获得PSI复合物。证明了PSI异质性的存在。 相似文献
19.
高分子量(HMW)DNA的制备是进行生物基因组物理作图等相关基因组学研究至关重要的基础工作。为建立一种高效的提取大麦高分子量DNA的方法,对琼脂糖包埋原生质体和琼脂糖包埋细胞核分离基因组DNA的方法进行了改良和优化。限制酶消化,脉冲电场凝胶电泳(PFGE)和Southern杂交等检测表明,应用优化的实验方法所分离的大麦基因组HMW DNA质量好,数量多。与原方法相比,新改进的方法实验流程缩短,效率提高,所分离HMWDNA能更好地满足后续相关研究的需要。这两种优化的分离大麦HMW DNA技术亦可应用于其它禾谷类植物基因组DNA的制备。 相似文献