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1.
《Food Control》2015
Bacillus cereus can cause emetic and diarrheal food poisoning. It is widespread in nature and therefore, considered a major foodborne pathogen. To develop a sensitive and reliable assay for detecting enterotoxin genes (nheA, entFM, hblD, cytK) and emetic toxin (ces), specific primers each targeting one individual gene were designed. Propidium monoazide (PMA) was coupled with the developed multiplex PCR (mPCR) for the detection of viable B. cereus. The inclusivity and exclusivity of the PMA-mPCR was confirmed using a panel of 44 strains including 17 emetic and 9 enterotoxic B. cereus reference strains and 18 non-target strains. The limit of detection (LOD) without PMA treatment in pure DNA was 2 pg/reaction tube. The LOD of mPCR assay in pure heat-killed dead bacteria was 4.0 × 102 CFU/mL. Also, the LOD on the viable bacteria with or without PMA treatment was similar (3.8 × 102 CFU/mL) showing that the PMA treatment did not significantly decrease sensitivity. Finally, the newly developed PMA-mPCR successfully detected 4.8 × 103 and 3.6 × 103 CFU/g of viable B. cereus F4810/72 (emetic) and B. cereus ATCC 12480 (enterotoxic) reference strains, respectively, in food samples. Hence, this study combines PMA and mPCR to detect viable B. cereus with a wide range of toxin detection (5 toxins). Thus, the novel PMA-mPCR assay developed in this study is a rapid and efficient diagnostic tool for the monitoring of viable B. cereus in food samples and potentially other samples via appropriate DNA extraction. 相似文献
2.
《Food Control》2015
A diverse range of genetic elements has been used to develop genetically modified organisms (GMOs) over the last 18 years. Screening methods that target few elements, such as the Cauliflower Mosaic Virus 35S promoter (P-35S) and Agrobacterium tumefaciens nopaline terminator (T-nos), are not sufficient to screen GMOs. In the present study, a multiplex PCR system for all globally commercialized GM soybean events was developed to easily trace the events. For this purpose, screening elements of 24 GM soybean events were investigated and 9 screening targets were selected and divided into three individual triplex PCR systems: P-35S, ribulose-1,5-bisphosphate carboxylase small subunit promoter of Arabidopsis thaliana, T-nos, T-35S, pea E9 terminator, open reading frame 23 terminator of A. tumefaciens, proteinase inhibitor II terminator of potato, acetohydroxy acid synthase large subunit terminator of A. thaliana, and the revealed 3′ flanking sequences of DP-305423-1. The specificity of the assays was confirmed using thirteen GM soybean events as the respective positive/negative controls. The limit of detection of each multiplex set, as determined using certified reference materials of specific GM events, ranged from 0.03 to 0.5%, depending upon target. Furthermore, 26 food samples that contained soybean ingredients, which were purchased from the USA, China, Japan, and Korea, were analyzed, 17 of which contained one or more GM soybean events. These results suggest that the developed screening method can be used to efficiently track and identify 24 GM soybean events in food and feed. 相似文献
3.
Maria Grazia Basanisi Gianfranco La Bella Gaia Nobili Rosa Coppola Annita Maria Damato Maria Assunta Cafiero Giovanna La Salandra 《International Journal of Food Science & Technology》2020,55(3):1145-1150
The authenticity and traceability of meat products are issues of primary importance to ensure food safety. Unfortunately, food adulteration (e.g. the addition of inexpensive cuts to minced meat products) and mislabelling (e.g. the inclusion of meat from species other than those declared) happens frequently worldwide. The aim of this study was to apply a droplet digital PCR assay for the detection and quantification (copies μL−1) of the beef, pork, horse, sheep, chicken and turkey in meat products. The analysis conducted on commercial meat showed the presence of traces of DNA from other animal species than those declared. We show that the method is highly sensitive, specific and accurate (accuracy = 100%). This method could be adopted by competent food safety authorities to verify compliance with the labelling of meat products and to ensure quality and safety throughout the meat supply chain, from primary production to consumption. 相似文献
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文章简要阐述了PID码在多节目码流的复用与解复用中所起的作用。并运用PID码的原理来解释和解决在实际工作中碰到的PID码问题。 相似文献
7.
27例乙型血友病患者Ⅸ因子基因突变研究 总被引:1,自引:0,他引:1
采用PCR(聚合酶链反应)及GAWTS(GenomicAmplificationwithTranscriptsSequencing)技术,研究了江苏、湖北、山东、广东、福建、宁夏六省区27例乙型血友病患者及其家系成员FⅨ(九因子)基因各2.2Kb DNA序列。在2l例中发现20种不同类型的突变。其中12种是未曾报道的新突变。38%的突变发生在CpG二核苷致序列上,进一步证实了CpG确系突变热点。同时检出6例女性为致病基因携带者。对开展基因产前诊断及优生优育等,具有重要意义。 相似文献
8.
牙鲆淋巴囊肿病的PCR诊断方法研究 总被引:14,自引:0,他引:14
以中国养殖牙鲆 (Paralichthysolivaceus)淋巴囊肿病毒 (Lymphocystisdiseasevirus,LCDVcn)主要衣壳蛋白 (majorcapsidprotein ,MCP)基因的中间保守序列为目标基因 ,设计了一对特异性引物。该引物可扩增出 172bp的病毒DNA片段 ,其最小DNA检出量为 0 0 183ng。用PCR法从人工感染淋巴囊肿病毒 3天的牙鲆血、鳃、肝、脾、肠、胃及自然发病牙鲆的肿瘤中 ,分别检测到了LCDV的存在。本实验结果证明 ,PCR法对于早期检测LCDV是十分有效的。 相似文献
9.
Frédéric Debode Eric Janssen Gilbert Berben 《European Food Research and Technology》2007,226(1-2):273-280
In this paper, four different physical treatments (microwaves, heating by conduction, sonication and pressure autoclaving)
were performed to degrade a pure DNA extract, and their influence on GMO quantification was studied. The aim was to check
the hypothesis that processing of agrofood products results in a similar degradation rate for both the transgenic target and
the specific target. Indeed we could observe that even if the used physical treatments could lead to a significant increase
of C
t values for both transgenic and specific targets, the resulting ΔC
t remained stable. So, the main conclusion of the study is that the aforementioned hypothesis seems valid and thus a physical
degradation of DNA will not affect the relative quantitation of the GMO content, provided that both the specific and transgenic
targets have very similar size. A second important issue of the experiments performed was that DNA is a very robust molecule
as it is extremely difficult to reach a mean size below 100 bp. The study also gives evidence of the importance of using small
targets. 相似文献
10.
转基因食品安全性的商榷 总被引:2,自引:0,他引:2
转基因生物技术近年来发展非常迅速,被广泛应用于食品领域,转基因食品的安全性在全球范范围内引了激烈的争论。转基因食品被越来越多的国家要求贴示标签,相应的法规相继出台。不论转基因食品安全与否,对其的检测技术都是必不可少的。 相似文献