為了紀念科學引文索引（SCI）誕生五十周年，Nature委托湯森路透列出了數據庫中的百大高引論文。榮登的那篇文章你一定猜不到。
據介紹，這份名單的入圍資格是引用次數超過12,119。令人意外的是，許多特別的論文并未入榜，比如DNA雙螺旋結構的確定。而1951年發表的一種蛋白定量方法，狂攬三十多萬次引用，成為了zui大的贏家。該文章的作者，美國生化學家Oliver Lowry也對此感到困惑。
| Rank | Title | Times cited |
| 1 | Protein measurement with the folin phenol reagent. | 305148 |
| 2 | Cleavage of structural proteins during the assembly of the head of bacteriophage T4. | 213005 |
| 3 | A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. | 155530 |
| 4 | DNA sequencing with chain-terminating inhibitors. | 65335 |
| 5 | Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. | 60397 |
| 6 | Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. | 53349 |
| 7 | Development of the Colle-Salvetti correlation-energy formula into a functional of the electron density. | 46702 |
| 8 | Density-functional thermochemistry. III. The role of exact exchange. | 46145 |
| 9 | A simple method for the isolation and purification of total lipides from animal tissues. | 45131 |
| 10 | Clustal W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. | 40289 |
| 11 | Nonparametric estimation from incomplete observations. | 38600 |
| 12 | Basic local alignment search tool. | 38380 |
| 13 | A short history of SHELX. | 37978 |
| 14 | Gapped BLAST and PSI-BLAST: A new generation of protein database search programs. | 36410 |
| 15 | A revised medium for rapid growth and bio assays with tobacco tissue cultures. | 36132 |
| 16 | Generalized gradient approximation made simple. | 35405 |
| 17 | "Mini-mental state": A practical method for grading cognitive state of patients for clinician. | 34532 |
| 18 | A rapid method of total lipid extraction and purification. | 32131 |
| 19 | Detection of specific sequences among DNA fragments separated by gel-electrophoresis. | 31904 |
| 20 | The neighbor-joining method: A new method for reconstructing phylogenetic trees. | 30176 |
| 21 | Analysis of relative gene expression data using real-time quantitative PCR and the 2(T)(-Delta Delta C) method. | 28870 |
| 22 | Revised effective ionic radii and systematic studies of interatomic distances in halides and chalcogenides. | 28658 |
| 23 | Processing of X-ray diffraction data collected in oscillation mode. | 28647 |
| 24 | Regression models and life-tables. | 28439 |
| 25 | Density-functional exchange-energy approximation with correct asymptotic-behavior. | 26475 |
| 26 | Colorimetric method for determination of sugars and related substances. | 25735 |
| 27 | Use of lead citrate at high pH as an electron-opaque stain in electron microscopy. | 24449 |
| 28 | The CLUSTAL_X Windows interface: Flexible strategies for multiple sequence alignment aided by quality analysis tools. | 24098 |
| 29 | Statistical methods for assessing agreement between two methods of clinical measurement. | 23826 |
| 30 | Reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. | 23642 |
| 31 | Isolation of biologically-active ribonucleic-acid from sources enriched in ribonuclease. | 23435 |
| 32 | The attractions of proteins for small molecules and ions. | 23421 |
| 33 | The moderator–mediator variable distinction in social psychological-research — conceptual, strategic, and statistical considerations. | 23356 |
| 34 | Self-consistent equations including exchange and correlation effects. | 23059 |
| 35 | Rapid colorimetric assay for cellular growth and survival — application to proliferation and cyto-toxicity assays. | 23011 |
| 36 | Helical microtubules of graphitic carbon. | 22899 |
| 37 | The colorimetric determination of phosphorus. | 22690 |
| 38 | Disc electrophoresis — II. Method and application to human serum proteins. | 22074 |
| 39 | Inhomogeneous electron gas. | 21931 |
| 40 | A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. | 21446 |
| 41 | Confidence limits on phylogenies: an approach using the bootstrap | 21373 |
| 42 | A new generation of Ca2+ indicators with greatly improved fluorescence properties | 19561 |
| 43 | Efficient iterative schemes for ab initio total-energy calculations using a plane-wave basis set. | 18856 |
| 44 | High-resolution 2-dimensional electrophoresis of proteins. | 18489 |
| 45 | MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. | 18286 |
| 46 | Fuzzy sets. | 18203 |
| 47 | Phase annealing in SHELX-90: direct methods for larger structures. | 17728 |
| 48 | Clinical diagnosis of Alzheimer’s disease: Report of the NINCDS-ADRDA Work Group under the auspices of Department of Health and Human Services Task Force on Alzheimer’s Disease. | 17220 |
| 49 | Special points for Brillouin-zone integrations. | 17087 |
| 50 | Study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. | 17067 |
| 51 | The CES-D scale: a self-report depression scale for research in the general population. | 17055 |
| 52 | Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. | 17025 |
| 53 | A rating scale for depression. | 16734 |
| 54 | An inventory for measuring depression. | 16264 |
| 55 | A simple method for displaying the hydropathic character of a protein. | 16059 |
| 56 | Determination of serum proteins by means of the biuret reaction. | 16009 |
| 57 | Maximum likelihood from incomplete data via EM algorithm. | 15993 |
| 58 | Equation of state calculations by fast computing machines. | 15902 |
| 59 | Controlling the false discovery rate: a practical and powerful approach to multiple testing. | 15898 |
| 60 | Measurement of protein using bicinchoninic acid. | 15802 |
| 61 | The assessment and analysis of handedness: the Edinburgh inventory. | 15517 |
| 62 | Estimation of concentration of low-density lipoprotein cholesterol in plasma, without use of preparative ultracentrifuge. | 15469 |
| 63 | Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. | 15160 |
| 64 | Multiple range and multiple F tests. | 15047 |
| 65 | Electric field effect in atomically thin carbon films. | 15022 |
| 66 | Tissue sulfhydryl groups. | 15019 |
| 67 | Isolation of mononuclear cells and granulocytes from human blood. | 14934 |
| 68 | The measurement of observer agreement for categorical data. | 14903 |
| 69 | Crystallography & NMR system: a new software suite for macromolecular structure determination. | 14898 |
| 70 | Gaussian-basis sets for use in correlated molecular calculations. 1. The atoms boron through neon and hydrogen. | 14617 |
| 71 | PROCHECK: a program to check the stereochemical quality of protein structures. | 14462 |
| 72 | The MOS 36-item short-form health survey (SF-36): I. Conceptual framework and item selection. | 14332 |
| 73 | A new look at statistical-model identification. | 14275 |
| 74 | Improved M13 phage cloning vectors and host strains — nucleotide-sequences of the M13mp18 and pUC19 vectors | 14232 |
| 75 | A comprehensive set of sequence-analysis programs for the vax. | 14226 |
| 76 | MODELTEST: Testing the model of DNA. | 14099 |
| 77 | From ultrasoft pseudopotentials to the projector augmented-wave method. | 14049 |
| 78 | Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. | 13881 |
| 79 | Comparison of simple potential functions for simulating liquid water. | 13774 |
| 80 | The development and use of quantum-mechanical molecular-models. 76. AM1: a new general-purpose quantum-mechanical molecular-model | 13718 |
| 81 | Phosphorus assay in column chromatography. | 13523 |
| 82 | MOLSCRIPT: a program to produce both detailed and schematic plots of protein structures. | 13496 |
| 83 | Van der Waals volumes and radii. | 13417 |
| 84 | A new and rapid colorimetric determination of acetylcholinesterase activity. | 13332 |
| 85 | Projector augmented-wave method. | 13330 |
| 86 | Optimization by simulated annealing. | 13293 |
| 87 | Nitric oxide: physiology, pathophysiology, and pharmacology. | 13267 |
| 88 | An algorithm for least-squares estimation of nonlinear parameters. | 13258 |
| 89 | Efficiency of ab initio total energy calculations for metals and semiconductors using a plane-wave basis set. | 13084 |
| 90 | A low-cost, high-efficiency solar-cell based on dye-sensitized colloidal TiO2 films. | 12873 |
| 91 | A low-viscosity epoxy resin embedding medium for electron microscopy. | 12807 |
| 92 | The Protein Data Bank. | 12754 |
| 93 | Accurate and simple analytic representation of the electron-gas correlation-energy | 12748 |
| 94 | Rapid alkaline extraction procedure for screening recombinant plasmid DNA. | 12721 |
| 95 | Improved methods for building protein models in electron-density maps and the location of errors in these models. | 12649 |
| 96 | Accurate spin-dependent electron liquid correlation energies for local spin-density calculations — a critical analysis. | 12583 |
| 97 | Continuous cultures of fused cells secreting antibody of predefined specificity. | 12391 |
| 98 | Homeostasis model assessment: insulin resistance and beta-cell function from fasting plasma glucose and insulin concentrations in man. | 12257 |
| 99 | Adsorption of gases in multimolecular layers. | 12252 |
| 100 | MrBayes 3: Bayesian phylogenetic inference under mixed models | 12209 |
| 101 | Atherosclerosis — an inflammatory disease. | 12119 |
那么榜上還有哪些與生物學有關的關鍵方法呢？
生物學技術
引文主要被蛋白質生化論文所占據。1951年發表的Lowry蛋白定量法遙遙，而Bradford蛋白定量法位列第三。排在它們之間的是用于蛋白分析的Laemmli緩沖液。這些技術在細胞和分子生物學文章中被大量引用，它們仍是這些領域zui基本的工具之一。
上榜的生物學技術中有兩個獲得了諾貝爾獎。Frederick Sanger的DNA測序方法位列第四，該方法也為他贏得了1980年的諾貝爾化學獎。Kary Mullis因PCR法獲得了1993年的諾貝爾化學獎，這一方法在引文中位列63。測序和PCR都是遺傳學研究的革命性技術，如今仍然有著旺盛的生命力。
除此之外，上世紀八十年代的一種RNA純化技術位列第五。該技術實現了低成本的快速RNA提取，受到了廣泛的歡迎。在非編碼短RNA的研究熱潮中，這一技術仍然起著重要的作用。
生物信息學
基因測序的迅猛發展，大大提高了序列分析論文的排名，BLAST就是一個很好的例子。對于研究基因和蛋白功能的研究者來說，BLAST近二十年來算得上是*。我們只需要打開一個網頁，輸入DNA、RNA或蛋白序列，就能在幾秒鐘內獲得各種生物中的相關序列。BLAST的兩個版本居然都上了榜，分別在12位和14位。
能夠一次比對多個序列的Clustal也不甘示弱，ClustalW和ClustalX的排名分別是第10和第28。在Clustal軟件的幫助下，人們能研究源自不同生物的序列有怎樣的進化關系，看起來不想關的序列存在怎樣的匹配，預測DNA或蛋白特定位點發生改變對其功能有何影響。
系統發生
系統發生學主要是研究物種之間的進化關系，這類方法也隨著基因組測序的發展而水漲船高。
榜的第二十位是neighbor-joining法。這種方法由根井正利（Masatoshi Nei）等人提出，可以根據大量物種之間的進化距離，快速有效的構建進化樹。
排在第41位的文章，將統計學工具用到了系統發生研究中。這種稱為bootstrap的工具，能夠推斷進化樹不同部分的性。
榜上的其他生物學相關方法，還包括一些用在生物醫學領域的統計學方法，以及晶體結構分析用到的工具。（除了湯森路透的榜以外，Google學術搜索也排出了自己的名單，感興趣的同志們請點這里 ）
“如果你想要的是被引用，”耶魯大學的化學家Peter Moore評論道。“那么開發一種人們需要的方法比探尋宇宙的秘密更容易達到目的。”
原文檢索：
Richard Van Noorden,Brendan Maher & Regina Nuzzo. The 00 papers. Nature, 30 October 2014; doi:10.1038/514550a
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