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41.
Microsatellite DNA markers for rice chromosomes 总被引:45,自引:1,他引:44
H. Akagi Y. Yokozeki A. Inagaki T. Fujimura 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1996,93(7):1071-1077
We found 369 complete microsatellites, of which (CGG/GCC)n was the most frequent, in 11 798 rice sequences in the database. Of these microsatellites, 35 out of 45 could be successfully converted into microsatellite DNA markers using sequence information in their flanking regions. Thus, the time and labor used to develop new microsatellite DNA markers could be saved by using these published sequences. Twenty eight polymorphic markers between Asominori (japonica) and IR24 (indica) have been correctly mapped on the rice genome and microsatellites appear to be randomly distributed in the rice chromosomes. Integration of these markers with the published microsatellite DNA markers showed that about 35% of the rice chromosomes were covered by the 56 microsatellite DNA markers. These microsatellites were hypervariable and were easily to assay by PCR; they were distributed to all chromosomes and therefore, one can easily select plants carrying desired chromosome regions using these microsatellite DNA markers. Thus, microsatellite maps should aid the development of new breeds of rice saving time, labor, and money. 相似文献
42.
PCR analysis of oilseed rape cultivars (Brassica napus L. ssp. oleifera) using 5′ -anchored simple sequence repeat (SSR) primers 总被引:8,自引:0,他引:8
Y. M. Charters A. Robertson M. J. Wilkinson G. Ramsay 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1996,92(3-4):442-447
Primers complementary to simple sequence repeats (SSRs) and with variable three-base anchors at their 5 end, were used in PCR analyses to compare pooled DNA samples from various Brassica napus and B. rapa cultivars. Amplification products were resolved on polyacrylamide gels and detected by silver-nitrate staining. The resulting banding patterns were highly repeatable between replicate PCRs. Two of the primers produced polymorphisms at 33 and 23 band positions, respectively, and could each discriminate 16 of the 20 cultivars studied. Combined use of both primers allowed all 20 cultivars to be distinguished. The UPGMA dendrogram, based on the cultivar banding profiles, demonstrated clustering on the basis of winter/spring growth habit, high/low glucosinolate content, and cultivar origin (i.e. the breeder involved). Intracultivar polymorphism was investigated using a minimum of ten individuals for each cultivar and was found to vary considerably between cultivars. It is concluded that anchored SSR-PCR analysis is a highly informative and reproducible method for fingerprinting oilseed rape populations, but that intra-cultivar variation should be investigated before using banding profiles from pooled samples for the identification of individuals. 相似文献
43.
Identification of molecular markers in soybean comparing RFLP,RAPD and AFLP DNA mapping techniques 总被引:16,自引:0,他引:16
Jhy-Jhu Lin Jonathan Kuo Jin Ma James A. Saunders Hunter S. Beard Margaret H. MacDonald William Kenworthy George N. Ude Benjamin F. Matthews 《Plant Molecular Biology Reporter》1996,14(2):156-169
Three different DNA mapping techniques—RFLP, RAPD and AFLP—were used on identical soybean germplasm to compare their ability
to identify markers in the development of a genetic linkage map. Polymorphisms present in fourteen different soybean cultivars
were demonstrated using all three techniques. AFLP, a novel PCR-based technique, was able to identify multiple polymorphic
bands in a denaturing gel using 60 of 64 primer pairs tested. AFLP relies on primers designed in part on sequences for endonuclease
restriction sites and on three selective nucleotides. The 60 diagnostic primer pairs tested for AFLP analysis each distinguished
on average six polymorphic bands. Using specific primers designed for soybean fromEco RI andMse I restriction site sequences and three selective nucleotides, as many as 12 polymorphic bands per primer could be obtained
with AFLP techniques. Only 35% of the RAPD reactions identified a polymorphic band using the same soybean cultivars, and in
those positive reactions, typically only one or two polymorphic bands per gel were found. Identification of polymorphic bands
using RFLP techniques was the most cumbersome, because Southern blotting and probe hybridization were required. Over 50% of
the soybean RFLP probes examined failed to distinguish even a single polymorphic band, and the RFLP probes that did distinguish
polymorphic bands seldom identified more than one polymorphic band. We conclude that, among the three techniques tested, AFLP
is the most useful. 相似文献
44.
Joan E. Vickers Glenn C. Graham Robert J. Henry 《Plant Molecular Biology Reporter》1996,14(4):363-368
Amplification of thebar gene usingTaq DNA polymerase in PCR is often not successful, possibly due tobar's high GC content. We describe a PCR protocol in which reliable amplification at a sensitivity of one gene copy per genome
(in this study, barley) present in the reaction was achieved using a novel pair of primers and Expandtm High Fidelity DNA polymerase mix (Boehringer Mannheim). This method should allow for rapid screening of plants putatively
transformed withbar. 相似文献
45.
Gavin R. Sills William Bridges Salah M. Al-Janabi Bruno W. S. Sobral 《Molecular breeding : new strategies in plant improvement》1995,1(4):355-363
Saccharum robustum Brandes & Jesw. ex Grassl has been suggested as the immediate progenitor species of cultivated sugarcane (S. officinarum L.) [4]. Chromosome pairing and assortment in these two species were previously studied by genetic analysis of single-dose DNA markers in parents in and 44 F1 progeny of a cross between euploid, meiotically regular 2n=80S. officinarum LA Purple andS. robustum Mol 5829 [2]. This same population was subsequently clonally propagated and evaluated in replicated trials for quantitative traits important to sugarcane breeders. Numbers of stalks, tasseled stalks, and stalks with smut, and the average diameter of two stalks were determined one day prior to harvest. At harvest, plant material from each plot was weighed and evaluated for pol (sucrose content) and fiber percentages. Clones were significantly different (P<0.01) for all traits analyzed. Associations of 83 single-dose arbitrarily primed PCR genetic markers with quantitative trait loci (QTL) of recorded traits was determined by single-factor ANOVA, and multiple regression. QTL analysis revealed markers significantly (P<0.05) associated with the expression of each trait analyzed. Markers associated with QTL after multiple regression were tested for digenic linear × linear epistatic interactions. The various multilocus models explained between 23% and 58% of the total phenotypic variation and 32% and 76% of the genotypic variation for the various traits. Digenic interactions were uncommon. Implications for marker-assisted selection in sugarcane and sugarcane domestication are discussed. 相似文献
46.
Alec Breen Alan F. Rope Denise Taylor John C. Loper P. R. Sferra 《Journal of industrial microbiology & biotechnology》1995,14(1):10-16
Summary The use of DNA amplification fingerprinting (DAF) as a tool for monitoring mixed microbial populations in bioreactors was evaluated. Short (8-mer or 10-mer) oligonucleotides were used to prime DNA extracts from various biological reactors during polymerase chain reaction (PCR) amplification. The reactors examined in this study included two sets of anaerobic stirred tank continuous flow bioreactors. One set of anaerobic reactors was operated under methanogenic conditions and one set was operated under sulfate-reducing conditions. The anaerobic reactor communities in the methanol-fed reactors showed extensive DAF homology. DAF was also applied to a fixed-film azo dye degrading reactor to examine the degree of uniformity of colonization of the substratum in representative regions of the reactor. This method is a quick and relatively inexpensive means of monitoring microbial community structure during biological processes. Since no cultivation of the sample is involved, the genetic profile of the community is not biased by outgrowth conditions. DAF profiles may be useful for comparisons of population changes over time or of bench-scale vs pilot-scale reactors but not adequate for assessing community diversity. 相似文献
47.
陈清轩 《基因组蛋白质组与生物信息学报(英文版)》1995,(1)
DetectionandAnalysisofanEstrous-associatedOviductalGlycoproteinDNAFragmentfromPrimatesbyPCR¥CHENQing-xuan(陈清轩);ClaytonE.Walto... 相似文献
48.
49.
50.
采用聚合酶链反应(Polymeranse Chian Reation,即PCR)技术检测结核分枝杆菌Mycobacterium tuberculosis,一年多来共检测了100例结核(肺、肾结核)患者的痰和尿液标本,结果PCR检出阳性率为81%,对照用储菌涂片抗酸染色法,阳性率为58%,用常规培养法阳性率为20%。而对50例非结核患者的痰和尿液标本的检测,PCR法仍有6%的阳性率,而用涂片或常规培 相似文献