全文获取类型
收费全文 | 306篇 |
免费 | 20篇 |
国内免费 | 36篇 |
出版年
2023年 | 2篇 |
2022年 | 9篇 |
2021年 | 4篇 |
2020年 | 11篇 |
2019年 | 6篇 |
2018年 | 8篇 |
2017年 | 11篇 |
2016年 | 12篇 |
2015年 | 12篇 |
2014年 | 10篇 |
2013年 | 14篇 |
2012年 | 17篇 |
2011年 | 27篇 |
2010年 | 18篇 |
2009年 | 22篇 |
2008年 | 22篇 |
2007年 | 20篇 |
2006年 | 16篇 |
2005年 | 22篇 |
2004年 | 15篇 |
2003年 | 18篇 |
2002年 | 9篇 |
2001年 | 5篇 |
2000年 | 3篇 |
1999年 | 4篇 |
1998年 | 6篇 |
1997年 | 5篇 |
1996年 | 5篇 |
1995年 | 5篇 |
1994年 | 3篇 |
1993年 | 3篇 |
1992年 | 1篇 |
1991年 | 2篇 |
1990年 | 3篇 |
1988年 | 1篇 |
1986年 | 1篇 |
1984年 | 3篇 |
1981年 | 1篇 |
1980年 | 1篇 |
1979年 | 1篇 |
1978年 | 1篇 |
1977年 | 1篇 |
1975年 | 1篇 |
1971年 | 1篇 |
排序方式: 共有362条查询结果,搜索用时 15 毫秒
301.
Metabolomics has developed into a major tool in functional genomics and plant systems biology. The various methods used for
metabolomic analysis will be discussed from the analytical methods back to the preanalytical phase and the biological experiment.
Particularly aspects of the preanalytical phase of the analysis is dealt with, including the risks of artefact formation with
the various commonly used solvents. Metabolomics is like a snap shot, and conclusions from dynamic systems must be drawn with
great care as demonstrated with a biosynthetic study of salicylate in Catharanthus roseus cell cultures. 相似文献
302.
涡虫是动物界最早出现两侧对称、三胚层、营自由生活的扁形动物,也是由水生向陆生过渡的重要类群,因而在动物系统演化中占有重要地位。由于涡虫再生能力极强,因此成为研究细胞分化与去分化 相似文献
303.
Somashekar D. Venkateshwaran G. Srividya C. Krishnanand Sambaiah K. Lokesh B.R. 《World journal of microbiology & biotechnology》2001,17(3):317-320
The efficacy of three extraction methods for determining the lipid and fatty acid composition of six fungal cultures was studied. The extraction methods were: chloroform/methanol (2:1), hexane/isopropanol (3:2) and Soxhlet extraction by using hexane. The total lipid and fatty acid composition varied in fungal cultures depending on the extraction conditions. Of the three methods, chloroform/methanol (2:1) was found to be the best for extraction of lipid and fatty acids from fungal cultures. 相似文献
304.
As a basis for the development of predictive mathematical models in systems biology and a quantitative understanding of cellular metabolism, reliable experimental data sets of intracellular metabolites are indispensable. A prerequisite for the acquisition of such data is the identification of a suitable sample preparation method. In this work, the extraction procedure for the simultaneous measurement of a wide range of intracellular metabolites from adherent mammalian cells in culture was optimized. A screening of several commonly used extraction protocols with Madin-Darby canine kidney (MDCK) cells found the methanol/chloroform (MeOH/CHCl(3)) and MeOH/Boil methods to be promising candidates for further analysis by anion-exchange chromatography. Both methods were optimized based on experimental design techniques with four response variables: Nucleotide Content, Energy Charge, Fructose 1,6-Bisphosphate content (F16bP), and Absorption at 280 nm. After data evaluation and with the help of desirability functions, an overall optimum for the extraction conditions was found. Using optimal settings, the extraction performances for MDCK and Vero cell cultivations of both methods were compared. Both methods extracted nearly the same absolute amounts of intracellular metabolites, suggesting that these methods are equal. However, recoveries for nucleotide diphosphates were significantly above 100% for both methods. This most likely was due to remaining nucleotide kinase activity during extraction. After combining individual steps of both methods, recoveries close to 100% for all metabolites could be reached. Absolute values of intracellular metabolites extracted with this modified method are comparable to the results of the two previously optimized methods, indicating a good extraction procedure according to the chosen response variables. 相似文献
305.
Optimization of extraction process of Glycyrrhiza glabra polysaccharides by response surface methodology 总被引:1,自引:0,他引:1
Experimental design was used to investigate the effect of three parameters (extraction time, extraction number and ratio of water to raw material) on polysaccharides yields. The ranges of the factors investigated were 3.5–4.5 h for extraction time (X1), 4–6 for extraction number (X2), and 25–35 for ratio of water to raw material (X3). The statistical analysis of the experiment indicated that extraction time and ratio of water to raw material had significant effect on Glycyrrhiza glabra polysaccharides yields. The central composite design showed that polynomial regression models were in good agreement with the experimental results with the coefficients of determination of 0.924 for Glycyrrhiza glabra polysaccharides yield. The optimal condition for Glycyrrhiza glabra polysaccharides yield within the experimental range of the variables studied was at 4.3 h, 6, and 35. At this condition, the predicted yield of polysaccharides extracted was 3.6%. 相似文献
306.
Osytek A Biesaga M Pyrzynska K Szewczyńska M 《Journal of biochemical and biophysical methods》2008,70(6):1283-1286
Workers involved in the manufacture of drug substances may be exposed to active pharmaceuticals by inhalation of drug dusts or droplets which has been considered the main exposure route. The proposed HPLC method allowed to determine sulpiryde, hydroxyurea and dyprophylline in the concentration range of 0.01–0.187 mg/m3, 0.001–0.08 mg/m3 and 0.01–0.40 mg/m3 for sulpiryde, hydroxyurea and dyprophylline, respectively, when 480 L of air sample was collected on the glass fibre filters. Sulpiryde was extracted with a solvent system consisting of acetonitrile–phosphate buffer at pH 3 (85:15, v/v), while the best efficiency of extraction for hydroxyurea and dyprophylline was achieved using water. HPLC analysis of sulpiryde with fluorescence detection was more sensitive (LOD = 3.1 μg/L) in comparison with UV detection (LOD = 84.4 μg/L). 相似文献
307.
工业化废水处理反应器污泥总DNA提取方法 总被引:2,自引:0,他引:2
根据工业化废水处理反应器污泥特性,对常规的溶菌酶-SDS-酚/氯仿环境样品总DNA提取方法进行改进,增强样品预处理,强化细胞裂解,提高杂质去除效率,获得了一种工业化污泥总DNA提取的通用方法,并采用该方法对石家庄若干实际运行的工业化厌氧、好氧反应器的污泥样品进行了总DNA提取研究.结果表明,该方法对所选污泥样品均有效,具有普适性.提取的污泥总DNA杂质含量少,纯度高,A260/A280在1.8左右;提取效率较高.总DNA产率都在0.7 mg/g以上,最大产率可达0.85 mg/g.所提取的污泥总DNA可以直接作为模板进行PCR反应,PCR产物直接进行变性梯度凝胶电泳(DGGE),能够得到较好的DGGE谱图,表明该方法提取的污泥总DNA样品可满足后续分析研究的要求. 相似文献
308.
一种改进的丝状真菌DNA提取方法 总被引:10,自引:0,他引:10
以丝状真菌雅致枝霉(Thamnidium elegans)和深黄伞形霉(Umbelopsis isabellina)为材料,使用优化的CTAB法提取基因组DNA.改进后的方法使用液氮冻融以及玻璃珠振荡的方法代替了传统的液氮研磨,所需茵体量少,而得到的基因组DNA比用传统的CTAB法得到的基因组DNA产率高,纯度好、且步骤简单,适用于一次微量提取多个样品的基因组DNA.这种方法得到的基因组DNA可用于大部分分子生物学基本实验如PCR和DNA的酶切等. 相似文献
309.
土壤溶解性有机质的特性与环境意义 总被引:38,自引:4,他引:34
土壤生态环境是一个复杂的多介质多界面体系,尽管关于土壤溶解性有机质 研究还不完善,但现有的研究表明,它是这一环境中最为活跃的化学组成之一。由于土壤溶解性有机质在C、N、P和S等营养元素的生物地球化学过程、成土过程、微生物的生长代谢过程、土壤有南分解和转化过程以及土壤污染物的迁移过程有着重要的作用,因此已成为土壤科学、生态科学和环境科学交叉领域的研究热点,本文从土壤溶解性有机质的提取方法、来源、组成、含量和影响因素、生物有效性及环境意义等方面的研究进展作了简要的论述,同时提出了未来的研究方向。 相似文献
310.