全文获取类型
收费全文 | 10915篇 |
免费 | 797篇 |
国内免费 | 439篇 |
专业分类
12151篇 |
出版年
2024年 | 50篇 |
2023年 | 171篇 |
2022年 | 175篇 |
2021年 | 199篇 |
2020年 | 355篇 |
2019年 | 373篇 |
2018年 | 377篇 |
2017年 | 332篇 |
2016年 | 334篇 |
2015年 | 283篇 |
2014年 | 763篇 |
2013年 | 1082篇 |
2012年 | 609篇 |
2011年 | 529篇 |
2010年 | 368篇 |
2009年 | 460篇 |
2008年 | 485篇 |
2007年 | 555篇 |
2006年 | 435篇 |
2005年 | 475篇 |
2004年 | 336篇 |
2003年 | 326篇 |
2002年 | 268篇 |
2001年 | 271篇 |
2000年 | 194篇 |
1999年 | 203篇 |
1998年 | 222篇 |
1997年 | 170篇 |
1996年 | 189篇 |
1995年 | 243篇 |
1994年 | 228篇 |
1993年 | 189篇 |
1992年 | 169篇 |
1991年 | 112篇 |
1990年 | 82篇 |
1989年 | 58篇 |
1988年 | 53篇 |
1987年 | 35篇 |
1986年 | 48篇 |
1985年 | 46篇 |
1984年 | 53篇 |
1983年 | 42篇 |
1982年 | 43篇 |
1981年 | 36篇 |
1980年 | 36篇 |
1979年 | 24篇 |
1978年 | 14篇 |
1977年 | 13篇 |
1976年 | 12篇 |
1973年 | 8篇 |
排序方式: 共有10000条查询结果,搜索用时 0 毫秒
101.
Cobalt determinations in biological fluids are of great interest in biological or toxicological research programs. Cobalturia
is often chosen as an indicator for a biological monitoring program in occupational exposure to cobalt dusts. The method described
here derives from the IUPAC reference method for nickel determination. It enables cobaltemia and cobalturia to be measured
in small samples (1 mL). The mean usual values for cobalt in biological fluids are very low (2.7 nmol L−1 for serum and 6.7 nmol L−1 for urine), and therefore, thus require an analytical procedure with preconcentration and extraction. The sample is mineralized
by wet acid digestion. After digestion, inorganic cobalt is extracted in form of ammonium pyrrolidine dithiocarbamate complex
into isobutyl methyl ketone and measured in the organic layer by electrothermal atomic absorption spectrometry.
The analytical parameters are described in detail. The extraction output is about 99%. The detection limits are 1.93 and 1.89
nmol L−1 for serum and urine, respectively. Sensitivity (expressed as the concentration that gives a 0.044 absorbance) is 3.4 nmol
L−1 for serum and 3.3 nmol L−1 for urine.
Within-run precision ranged between 3.9 and 2.5% (coefficients of variation) for serum and 4.2 and 1.1% for urine, at 87 and
136 nmol L−1 levels, respectively. Between-run precision ranged between 4.3 and 3.3% (coefficients of variation) for serum and 4.2 and
2.3% for urine, at 87 and 136 nmol L−1 levels, respectively. At very low concentration, 5.7 nmol L−1 for serum and 2.5 nmol L−1 for urine, the between-run precision is, respectively, 19.5 and 28%.
Linearity is effective between 0 and 272 nmol L−1. Interferences and matrix effects are negligible for urine, serum, or plasma samples without hemoglobin. The method is easily
applicable for routine determinations. 相似文献
102.
George N. Rudenko Caius M. T. Rommens H. John J. Nijkamp Jacques Hille 《Plant molecular biology》1993,21(4):723-728
We describe a novel modification of the polymerase chain reaction for efficient in vitro amplification of genomic DNA sequences flanking short stretches of known sequence. The technique utilizes a target enrichment step, based on the selective isolation of biotinylated fragments from the bulk of genomic DNA on streptavidin-containing support. Subsequently, following ligation with a second universal linker primer, the selected fragments can be amplified to amounts suitable for further molecular studies. The procedure has been applied to recover T-DNA flanking sequences in transgenic tomato plants which could subsequently be used to assign the positions of T-DNA to the molecular map of tomato. The method called supported PCR (sPCR) is a simple and efficient alternative to techniques used in the isolation of specific sequences flanking a known DNA segment. 相似文献
103.
Dr Sachio Hayashi Sinji Sasao Yoshiyuki Takasaki Kiyohisa Imada 《Journal of industrial microbiology & biotechnology》1994,13(2):103-105
Summary -Fructofuranosidase, which produces fructo-oligosaccharides (1-kestose and nystose) from sucrose, was purified fromAureobasidium and immobilized on DEAE-cellulose at especially high efficiency (95%). The enzymatic profiles of the immobilized enzyme were almost identical to those of the native form except that the stability was slightly improved. The immobilized enzyme was stable during long-term continuous reaction for up to 360 h. 相似文献
104.
105.
本文研究了超声波和相转移催化剂在Reimer—Tiemann反应中的应用,提出了超声波和相转移催化下Reimer—Tiemann反应的机理。研究结果表明:在超声波和相转移催化剂的共同作用下,二氯卡宾形成十分迅速,羟基苯甲醛产率显著提高,反应时间成倍缩短。 相似文献
106.
J. J. Windig 《Journal of evolutionary biology》1994,7(6):665-695
The genetic basis of the dry-wet season polyphenism of wing pattern in response to temperature shown by Bicyclus anynana was studied, using a split-family design over four temperatures. Reaction norms crossed, but were only linear in the three highest temperatures, and only when larval development time was used as the environmental axis. Significant full-sib additive variances (VA) and heritabilities (h2) for plasticity were found using slopes of reaction norms in a bootstrap procedure. Heritabilities were lower in intermediate temperatures, mainly due to differences in the residual variances (VR). There was no clear trend in VA across temperatures, contrary to the expectation that VA would have been depleted by natural selection at the extreme temperatures and not depleted at the intermediate temperatures which occur less frequently in the field. Unpredictability in the onset of the following season at intermediate temperatures might lead to selection for diverse flresponses resulting in relatively high VRs. Theoretical models linking reaction norms to genetic parameters in separate environments were difficult to apply in this study, particularly because they are based on the assumption that VRs are constant. However the reaction norm approach combined with quantitative genetics provided a valuable insight into the evolution of the observed polyphenism. 相似文献
107.
Ralph Rapley 《Molecular biotechnology》1994,2(3):295-298
The polymerase chain reaction (PCR) is a powerful core molecular biology technique, which when coupled to chain termination
sequencing allows gene and DNA sequence information to be derived rapidly. A number of modifications to the basic PCR format
have been developed in an attempt to increase amplification efficiency and the specificity of the reaction. We have applied
the use of DNA-binding protein, gene 32 protein from bacteriophage T4 (T4gp32) to increase amplification efficiency with a
number of diverse templates. In addition, we have found that using single-stranded DNA-binding protein (SSB) or recA protein
in DNA sequencing reactions dramatically increases the resolution of sequencing runs. The use of DNA-binding proteins in amplification
and sequencing may prove to be generally applicable in improving the yield and quality of a number of templates from various
sources. 相似文献
108.
A polymerase chain reaction (PCR) method for discriminating Meloidogyne incognita, M. arenaria, M. javanica, M. hapla, and M. chitwoodi was developed. Single juveniles were ruptured in a drop of water and added directly to a PCR reaction mixture in a microcentrifuge tube. Primer annealing sites were located in the 3'' portion of the mitochondrial gene coding for cytochrome oxidase subunit II and in the 16S rRNA gene. Following PCR amplification, fragments of three sizes were detected. The M. incognita and M. javanica reactions produced a 1.7-kb fragment; the M. arenaria reaction, a 1.1-kb fragment; and the M. hapla and M. chitwoodi reactions resulted in a 0.52-kb fragment. Digestion of the amplified product with restriction endonucleases allowed discrimination among species with identically sized amplification products. Dra I digestions of the 0.52-kb amplification product produced a characteristic three-banded pattern in M. chitwoodi, versus a two-banded pattern in M. hapla. Hinf I digestion of the 1.7-kb fragment produced a two-banded pattern in M. javanica, versus a three-banded pattern in M. incognita. Amplification and digestion of DNA from juveniles from single isolates of M. marylandi, M. naasi, and M. nataliei indicated that the diagnostic application of this primer set may extend to less frequently encountered Meloidogyne species. 相似文献
109.
Application of computer to monitoring and control of fermentation process: Microbial conversion of ML-236B Na to pravastatin 总被引:1,自引:0,他引:1
An automatic feeding process for microbial hydroxylation of ML236B sodium salt (ML-236B Na; compactin) by Streptomyces carbophilus SANK 62585 was developed. The hydroxylated product, pravastatin sodium salt (pravastatin; trade name Mevalotin), is an inhibitor of 3-hydroxy-3-methyglutaryl-coenzyme A reductase (HMG-CoA reductase) used as cholesterol-lowering drug. The hydroxylation activity of S. carbophilus was induced by the addition of ML236B Na to culture broth but inhibited by high concentration of ML236B Na. In order to obtain high conversion yield, it was necessary to maintain optimum ML236B Na concentration throughout the fermentation by continuous feeding. For this purpose, we developed an on-line monitoring method, which mainly consisted of a cross-flow filtration module, high-performance liquid chromatography (HPLC) analyzer, feed pump, and microcomputer for regulation of ML236B Na concentration. An algorithm for control of ML236B Na feed rate based on feedback and feed-forward control where conversion rate after Deltat was estimated by using regression analysis of the five latest values of conversion rate. In a fed-batch culture employing this system, the concentration of ML236B Na was maintained at optimum level during the fermentation and the productivity of pravastatin was increased threefold over that obtained in manual control culture. (c) 1993 John Wiley & Sons, Inc. 相似文献
110.
This article demonstrates the successful in situ real-time monitoring of the cell concentration of Perilla frutescens in a bioreactor by using a laser turbidimeter. It was found that turbidity measurements at 780 nm with the laser sensor were hardly affected by the red color of the anthocyanin produced by P. frutescens cells, nor by the aeration rate or agitation speed within the ranges investigated. There was an excellent linear relationship, with a correlation coefficient (r(2)) higher than 0.99, between the sensor's response and the cell concentration. The whole growth stage of the cells, i.e., lag, logarithmic, and stationary phases, in bioreactor cultivations, could be satisfactorily estimated on-line by means of the in situ turbidimeter. However, during the declining phase of the cells, an apparent deviation was observed between the on-line estimations and off-line measurements of cell concentrations by dry cell weight, while the wet cell weight could be estimated by the same turbidimeter system. We found that this deviation was caused by a decrease in the cell density due to an increase of the individual cell volume and a decrease of the cell dry weight during the declining phase. (c) 1993 John Wiley & Sons, Inc. 相似文献