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1.
Synthesis of nucleic acid probes on membrane supports: a procedure for the removal of unincorporated precursors 总被引:1,自引:0,他引:1
S P Bhat 《Analytical biochemistry》1990,184(1):59-62
We have used DNA bound to small pieces of nylon membrane for the synthesis of radioactive probes. The DNA to be used for generating the probe(s) is first bound to nylon membranes and then introduced into the reaction mix. The labeling reaction takes place on the membrane and therefore allows easy removal of unincorporated precursors by simple washing for 1-2 min. The clean labeled probe is eluted from the membrane in formamide or in water and is ready for use. This DNA-membrane can be stored for reuse. Synthesis of probes on a solid support such as nylon membrane thus circumvents problems associated with chromatographic manipulations needed for the separation of labeled DNA from unicorporated precursors. Probes synthesized in this manner are as efficient in detecting nucleic acid sequences as those synthesized in solution. 相似文献
2.
E. Rai S. Sharma A. Koul A. K. Bhat A. J. S. Bhanwer R. N. K. Bamezai 《Human genetics》2008,123(1):117
3.
Anwesha Dutta Premalatha Shetty Smitha Bhat Yeshaswini Ramachandra Shrinidhi Hegde 《Journal of biomolecular techniques》2012,23(4):128-135
A solvent system that extracts a maximum number of metabolites belonging to diverse chemical classes from complex biofluids, such as plasma, may offer useful inputs to understand the metabolic and physiological state of an individual. The present study compared seven solvent systems for extraction of metabolites from plasma. The extracts were analyzed by mass spectrometry (MS) and MS/MS (MS2) using a quadrupole time-of-flight liquid chromatography/MS system in positive and negative modes of ionization. Metabolites with molecular mass below 400 were identified using Human Metabolome Database MS2 and MS search interfaces. The acetone/isopropanol (2:1) system yielded promising results in positive ionization mode, as the maximum number of MS and MS2 features was detected in the extract. It was found to be superior in extraction of various classes of metabolites, especially organic acids, nucleosides and nucleoside derivatives, and heterocyclic molecules. Glycerophosphocholines in the mass range of 400–700 were found to be efficiently extracted by the methanol/chloroform/water (8:1:1) system. In negative mode as well, the maximum number of MS2 features was detected in methanol/chloroform/water and acetone/isopropanol extracts. The fingerprints of molecular features obtained in the negative and positive modes differed from each other to a significant extent. 相似文献
4.
5.
Audesh Bhat Anil Koul Ekta Rai Swarkar Sharma M. K. Dhar R. N. K. Bamezai 《Human genetics》2008,123(1):115
6.
Haploid plants were regenerated in vitro from unpollinated ovules of niger (Guizotia abyssinica (L. f.) (Cass.) on Murashige and Skoog nutrient medium (MS) supplemented with 10 μM naphthaleneacetic acid or 10 μM NAA +
1.5 μM kinetin and 30 g/l sucrose. Gamborg (B5) medium was the best for plant regeneration (in comparison with MS, Nitsch
and Nitsch (NN), and Chu (N6) media) from cultured ovules, and 6.66 and 7.33 ovules of JNC-6 and Ootacamund cultivars were
involved in direct plant regeneration on this medium. Matured ovules (ovules collected one day before anthesis or on the day
of anthesis) only responded to cultural regimes and involved in direct plantlet development. Cytological preparation of root
tips and chloroplast counts in the guard cells of leaf stomata of regenerated plants confirmed their haploid nature.
This text was submitted by the authors in English. 相似文献
7.
AbstractDesert plant species commonly use seed dormancy to prevent germination during unfavorable environmental conditions and thus increase the probability of seedling survival. Seed dormancy presents a challenge for restoration ecology, particularly in desert species for which our knowledge of dormancy regulation is limited. In the present study the effect of gibberellic acid (GA3) and potassium nitrate (KNO3) on seed dormancy release was investigated on eight Arabian desert species. Both treatments significantly enhanced the germination of most species tested. GA3 was more effective than KNO3 in enhancing germination percentage, reducing mean germination time and synchronizing the germination in most of the studied species. Light requirement during germination was species-specific, but in general the presence of light promoted germination more effectively when combined with KNO3 and GA3. The wide variation in dormancy and germination requirements among the tested species is indicative of distinct germination niches, which might assist their co-existence in similar habitat/environmental conditions. Seed pre-treatments that optimize germination in this habitat must therefore be assessed for individual species to improve the outcomes of ecological restoration. 相似文献
8.
Dipnarayan Saha Vajinder Kumar Shripad Ramachandra Bhat Ramamurthy Srinivasan 《Plant Molecular Biology Reporter》2011,29(2):265-277
Isolation and characterization of promoters are important in understanding gene regulation and genetic engineering of crop
plants. Earlier, a pentatricopeptide repeat protein (PPR) encoding gene (At2g39230), designated as Lateral Organ Junction (LOJ) gene, was identified through T-DNA promoter trapping in Arabidopsis thaliana. The upstream sequence of the LOJ gene conferred on the reporter gene a novel LOJ-specific expression. The present study was aimed at identifying and characterizing
the cis-regulatory motifs responsible for tissue-specific expression in the −673 and +90 bases upstream of the LOJ gene recognized as LOJ promoter. In silico analysis of the LOJ promoter revealed the presence of a few relevant regulatory motifs and a unique feature like AT-rich inverted repeat. Deletion
analysis of the LOJ promoter confirmed the presence of an enhancer-like element in the distal region (−673/−214), which stimulates a minimal
promoter-like sequence in the −424/−214 region in a position and orientation autonomous manner. The −136/+90 region of the
LOJ promoter was efficient in driving reporter gene expression in tissues like developing anthers and seeds of Arabidopsis. A positive regulation for the seed- and anther-specific expression module was contemplated within the 5′ untranslated region
of the LOJ gene. However, this function was repressed in the native context by the lateral organ junction-specific expression. The present
study has led to the identification of a novel lateral organ junction-specific element and an enhancer sequence in Arabidopsis with potential applications in plant genetic engineering. 相似文献
9.
Using antibodies to rat neural cell adhesion molecules (NCAM), we analyzed the NCAM of adult human brain. Various regions of the brain were analyzed quantitatively by Western blot. Grey matter showed four bands of NCAM with apparent molecular weights of 180,000, 170,000, 140,000 and 120,000. White matter showed one major band with an apparent Mr of 120,000 and a minor band of 180,000. Cerebellar grey matter contained mainly 170,000, 140,000 and 120,000, white cerebellar white matter had only 180,000 and 120,000 M1 NCAMS. Spinal cord showed mainly 120,000 Mr NCAM. Deglycosylation using N-glycanase resulted in 170,000, 160,000, 130,000 and 110,000 Mr proteins, suggesting that the four forms of human NCAM are derived from individual polypeptides. The presence of 170,000 M1 NCAM is unique to human brain. 相似文献
10.
Summary The rhizosphere microflora of arecanut palm under continuous application of organic manures and inorganic fertilizers was
studied. The nutrients applied are 100 g N, 40 g P2O5 and 140 g K2O/palm/year in the form of organics and inorganics. The application of organic manure increased the microbial population.
The increase in microbial population was observed between the rhizosphere samples collected at 0–30cm and 30–60 cm depths.
The surface cultivation of soil increased the microbial population.Trichoderma sp. andAspergillus sp. dominated in therhizosphere of arecanut palm.
Contribution No. 208. Central Plantation Crops Research Institute, Vittal-574243, Karnataka, India. 相似文献