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排序方式: 共有127条查询结果,搜索用时 312 毫秒
11.
Taylor DL Booth MG McFarland JW Herriott IC Lennon NJ Nusbaum C Marr TG 《Molecular ecology resources》2008,8(4):742-752
High throughput sequencing methods are widely used in analyses of microbial diversity, but are generally applied to small numbers of samples, which precludes characterization of patterns of microbial diversity across space and time. We have designed a primer-tagging approach that allows pooling and subsequent sorting of numerous samples, which is directed to amplification of a region spanning the nuclear ribosomal internal transcribed spacers and partial large subunit from fungi in environmental samples. To test the method for phylogenetic biases, we constructed a controlled mixture of four taxa representing the Chytridiomycota, Zygomycota, Ascomycota and Basidiomycota. Following cloning and colony restriction fragment length polymorphism analysis, we found no significant difference in representation in 19 of the 23 tested primers. We also generated a clone library from two soil DNA extracts using two primers for each extract and compared 456 clone sequences. Community diversity statistics and contingency table tests applied to counts of operational taxonomic units revealed that the two DNA extracts differed significantly, while the pairs of tagged primers from each extract were indistinguishable. Similar results were obtained using UniFrac phylogenetic comparisons. Together, these results suggest that the pig-tagged primers can be used to increase ecological inference in high throughput sequencing projects on fungi. 相似文献
12.
Storage of nematodes in soil at -15 C for 1 to 16 weeks greatly increased nematode recovery by a sugar-flotation-sieving procedure. One week of exposure to -15 C killed all nematodes except Pratylenchus zeae and Tylenchorhynchus claytoni which were recoverable in decreasing numbers up to 10 weeks by the Baermann funnel method. Optimum storage temperature for survival of most nematode species was 13 C. The numbers of Meloidogyne incognita, T. claytoni, Belonolaimus Iongicaudatus, and P. zeae recoverable by either extraction method remained constant or increased when stored at 13-24 C for 16 weeks. This was also true for Helicotylenchtts dihystera and Xiphinema americanum extracted by the Baermann funnel technique, whereas the numbers retrieved by the sugar-flotation-sieving method decreased slightly. All species except T. claytoni decreased appreciably in soil stored at 36 C. 相似文献
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Manuel Garber Michael C Zody Harindra M Arachchi Aaron Berlin Sante Gnerre Lisa M Green Niall Lennon Chad Nusbaum 《Genome biology》2009,10(6):R60-6
The most recent release of the finished human genome contains 260 euchromatic gaps (excluding chromosome Y). Recent work has
helped explain a large number of these unresolved regions as 'structural' in nature. Another class of gaps is likely to be
refractory to clone-based approaches, and cannot be approached in ways previously described. We present an approach for closing
these gaps using 454 sequencing. As a proof of principle, we closed all three remaining non-structural gaps in chromosome
15. 相似文献
15.
Athe M. N. Tsibris Bette Korber Ramy Arnaout Carsten Russ Chien-Chi Lo Thomas Leitner Brian Gaschen James Theiler Roger Paredes Zhaohui Su Michael D. Hughes Roy M. Gulick Wayne Greaves Eoin Coakley Charles Flexner Chad Nusbaum Daniel R. Kuritzkes 《PloS one》2009,4(5)
High-throughput sequencing platforms provide an approach for detecting rare HIV-1 variants and documenting more fully quasispecies diversity. We applied this technology to the V3 loop-coding region of env in samples collected from 4 chronically HIV-infected subjects in whom CCR5 antagonist (vicriviroc [VVC]) therapy failed. Between 25,000–140,000 amplified sequences were obtained per sample. Profound baseline V3 loop sequence heterogeneity existed; predicted CXCR4-using populations were identified in a largely CCR5-using population. The V3 loop forms associated with subsequent virologic failure, either through CXCR4 use or the emergence of high-level VVC resistance, were present as minor variants at 0.8–2.8% of baseline samples. Extreme, rapid shifts in population frequencies toward these forms occurred, and deep sequencing provided a detailed view of the rapid evolutionary impact of VVC selection. Greater V3 diversity was observed post-selection. This previously unreported degree of V3 loop sequence diversity has implications for viral pathogenesis, vaccine design, and the optimal use of HIV-1 CCR5 antagonists. 相似文献
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Niall J Lennon Robert E Lintner Scott Anderson Pablo Alvarez Andrew Barry William Brockman Riza Daza Rachel L Erlich Georgia Giannoukos Lisa Green Andrew Hollinger Cindi A Hoover David B Jaffe Frank Juhn Danielle McCarthy Danielle Perrin Karen Ponchner Taryn L Powers Kamran Rizzolo Dana Robbins Elizabeth Ryan Carsten Russ Todd Sparrow John Stalker Scott Steelman Michael Weiand Andrew Zimmer Matthew R Henn Chad Nusbaum Robert Nicol 《Genome biology》2010,11(2):1-9
We present an automated, high throughput library construction process for 454 technology. Sample handling errors and cross-contamination are minimized via end-to-end barcoding of plasticware, along with molecular DNA barcoding of constructs. Automation-friendly magnetic bead-based size selection and cleanup steps have been devised, eliminating major bottlenecks and significant sources of error. Using this methodology, one technician can create 96 sequence-ready 454 libraries in 2 days, a dramatic improvement over the standard method. 相似文献
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Asmaa Mamoune Michel Bahuau Yamina Hamel Valérie Serre Michele Pelosi Florence Habarou Marie-Ange Nguyen Morel Bertrand Boisson Sabrina Vergnaud Mai Thao Viou Luc Nonnenmacher Monique Piraud Patrick Nusbaum Joseph Vamecq Norma Romero Chris Ottolenghi Jean-Laurent Casanova Pascale de Lonlay 《PLoS genetics》2014,10(11)
Aldolase A deficiency has been reported as a rare cause of hemolytic anemia occasionally associated with myopathy. We identified a deleterious homozygous mutation in the ALDOA gene in 3 siblings with episodic rhabdomyolysis without hemolytic anemia. Myoglobinuria was always triggered by febrile illnesses. We show that the underlying mechanism involves an exacerbation of aldolase A deficiency at high temperatures that affected myoblasts but not erythrocytes. The aldolase A deficiency was rescued by arginine supplementation in vitro but not by glycerol, betaine or benzylhydantoin, three other known chaperones, suggesting that arginine-mediated rescue operated by a mechanism other than protein chaperoning. Lipid droplets accumulated in patient myoblasts relative to control and this was increased by cytokines, and reduced by dexamethasone. Our results expand the clinical spectrum of aldolase A deficiency to isolated temperature-dependent rhabdomyolysis, and suggest that thermolability may be tissue specific. We also propose a treatment for this severe disease.
Abstract Summary
Using recent technical advances involving exome analysis, we identified a new missense mutation in the ALDOA gene, encoding a key enzyme in the glycolytic pathway. The patients presented with severe recurrent rhabdomyolysis without hemolytic anemia. The decrease of aldolase A activity in myoblasts was enhanced at high temperature and could explain the fever-induced rhabdomyolysis. By contrast, enzyme thermolability was not found in erythrocytes, possibly accounting for the unusual clinical phenotype of the patients. Enzyme thermolability was rescued by arginine supplementation in vitro but not by other chaperone compounds. 相似文献20.