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产品介绍

微生物多样性测序,又名扩增子测序是对特定长度的PCR产物或者捕获的片段进行测序,主要包括16S rRNA测序、18S rRNA测序、ITS测序及目标区域扩增子测序等。采用第二代高通量测序平台测定的16S/18S /ITS某个高变区域的序列,来反映环境样品在细菌、真菌、古菌分类方面物种之间的差异,对研究海洋、土壤、肠道粪便等环境中的微生物构成有重要的指导作用。

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凌恩生物根据不同的研究目的和需求,扩增子测序可以细分为如下4个产品:

16S rRNA测序:16S rRNA为编码原核生物核糖体小亚基rRNADNA序列,主要进行细菌或古菌的多样性分析细菌核糖体RNA(rRNA)有三种类型:5S rRNA(120bp)16S rRNA(约1540bp)和23S rRNA(约2900bp)。5S rRNA基因序列较短,包含的遗传信息较少,不适于细菌种类的分析鉴定;23S rRNA基因的序列太长,且其碱基的突变率较高,不适于鉴定亲缘关系较远的细菌种类;16S rRNA普遍存在于原核细胞中,且含量较高、拷贝数较多(占细菌RNA总量的80%以上),便于获取模板,功能同源性高,遗传信息量适中,适于作为细菌多样性分析的标准。16S rRNA编码基因序列共有10个保守区和9个高可变区。

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18S rRNA测序:18S rRNA为编码真核生物核糖体小亚基rRNADNA序列,反映样品中真核生物之间的种类差异。与细菌多样性分析类似,在真核微生物中也有三类核糖体RNA(rRNA),包括5.8S rRNA18S rRNA28S rRNA18S rRNA基因是编码真核生物核糖体小亚基的DNA序列,其中既有保守区,也有可变区(V1-V9,没有V6区)。保守区域反映了生物物种间的亲缘关系,而可变区则能体现物种间的差异,适用于作种级及以上的分类标准。

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ITS测序:该类测序主要对环境微生物中的真菌多样性进行分析。包含两个区域:ITS1位于真核生物核糖体rDNA序列18S5.8S之间;ITS2位于真核生物核糖体rDNA序列5.8S28S之间。 ITS序列是内源转录间隔区(Internally Transcribed Spacer),位于真菌18S5.8S28S rRNA基因之间,分别为ITS 1ITS 2。在真菌中,5.8S18S28S rRNA基因具有较高的保守性,而ITS由于承受较小的自然选择压力,在进化过程中能够容忍更多的变异,在绝大多数真核生物中表现出极为广泛的序列多态性。同时,ITS的保守型表现为种内相对一致,种间差异较明显,能够反映出种属间,甚至菌株间的差异。并且ITS序列片段较小(ITS 1ITS 2长度分别为350 bp400 bp),易于分析,目前已被广泛用于真菌不同种属的系统发育分析。

微测4.png微测5.png

功能基因扩增子测序:根据客户的研究需求,基于功能基因/持家基因/靶基因等,对特定基因的PCR产物进行测序。在自然界的各类环境中,都有微生物的存在,它们在各自的“岗位”上都发挥着或大或小、或多或少的作用。有一些具有特殊功能的微生物,由于其作用的重要性或特殊性而受到人们的广泛关注,这些具有特殊功能的微生物叫做功能微生物,如氨氧化细菌、硫细菌、硝化细菌等。它们在分类学上有可能差异很大,但却具有相同或类似的基因使其能够发挥同样的作用。支配这些功能细菌发挥重要功能的基因被称为功能基因,如amoAdsrBnxrA。目前凌恩生物已收集GeneBank中所有已知功能基因的序列及其对应的种属信息,构建自己的功能基因数据库。


技术流程

NGS平台:

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分析流程

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NGS扩增子测序

送样要求


样本类型1:环境样本(请使用干冰或者冰袋运送)

1)土壤≥5g

2)粪便≥2g

样本类型2:DNA样品(请使用干冰或者冰袋运送)

1)DNA总量≥500ng

2)浓度≥5ng/ul



技术特色

  • 多种可选引物,针对不同微生物类型研究方向

  • 1对1项目服务,直接对接生信分析师,没有中间环节,沟通更高效

  • 多组学关联分析,可关联代谢组,蛋白组研究结果,全方面深化研究

  • 多种个性化分析方案,全方位满足研究需要,助力科研成功


分析示例

微生物多样性测序.png


部分凌恩客户文章

发表时间文章标题杂志名影响因子合作单位
2019Phage combination therapies for bacterial wilt disease in tomatoNature Biotechnology31.864南京农业大学
2023RIN enhances plant disease resistance via root exudate-mediated assembly of disease-suppressive rhizosphere microbiotaMolecular Plant27.5南京农业大学
2022Long-term efect of epigenetic modifcation in plant–microbe interactions: modifcation of DNA methylation induced by plant growth growth promoting bacteria mediates promotion processMicrobiome16.837南京农业大学
2022Antimicrobial PCMX facilitates the volatile fatty acids production during sludge anaerobic fermentation: Insights of the interactive principles, microbial metabolic profiles and adaptation mechanismsChemical Engineering Journal16.744河海大学
2022Enhanced removal of organic, nutrients, and PFCs in the iron-carbon micro-electrolysis constructed wetlands: mechanism and iron cycleChemical Engineering Journal16.744河海大学
2024From phyllosphere to insect cuticles: silkworms gather antifungal bacteria from mulberry leaves to battle fungal parasite attacksMicrobiome15.5中国科学院
2020Competition for iron drives phytopathogen control by natural rhizosphere microbiomesnature microbiology14.3南京农业大学
2022Diversity, function and assembly of the Trifolium repens L. root-associated microbiome under lead stressJournal of Hazardous Materials14.224东北农业大学
2022Toward emerging contaminants removal using acclimated activated sludge in the gravity-driven membrane filtration systemJournal of Hazardous Materials14.224哈尔滨工业大学
2024Impact of deoxynivalenol on rumen function production and health of dairy cows Insights from metabolomics and microbiota analysisJournal of Hazardous Materials13.6吉林农业大学
2024Remediation of neonicotinoid-contaminated soils using peanut shell biochar and composted chicken manure: Transformation mechanisms of geochemical fractionsJournal of Hazardous Materials13.6扬州大学
2024Remediation potential of an immobilized microbial consortium with corn straw as a carrier in polycyclic aromatic hydrocarbons contaminated soilJournal of Hazardous Materials13.6南京农业大学
2024Algicidal activity synchronized with nitrogen removal by actinomycetes: Algicidal mechanism, stress response of algal cells, denitrification performance, and indigenous bacterial community co-occurrenceJournal of Hazardous Materials13.6西安建筑科技大学
2024Distribution characteristics and microbial synergistic degradation potential of polyethylene and polypropylene in freshwater estuarine sedimentsJournal of Hazardous Materials13.6河海大学
2023Biochar-immobilized Bacillus megaterium enhances Cd immobilization in soil and promotes Brassica chinensis growthJournal of Hazardous Materials13.6山东大学
2022Insight into the role of biogenic manganese oxides-assisted gravity-driven membrane filtration systems toward emerging contaminants removalWater Research13.4哈尔滨工业大学
2022Sustainable restoration of anoxic freshwater using environmentally-compatible oxygen-carrying biochar: Performance and mechanismsWater Research13.4河海大学
2022Di-(2-ethylhexyl) phthalate exacerbated the toxicity of polystyrene nanoplastics through histological damage and intestinal microbiota dysbiosis in freshwater Micropterus salmoidesWater Research13.4华南农业大学
2022Combined remediation effects of biochar and organic fertilizer on immobilization and dissipation of neonicotinoids in soilsEnvironment International13.352扬州大学
2019Initial soil microbiome composition and functioning predetermine future plant healthScience Advances13.293南京农业大学
2023Potential negative effects of illumination on gravity-driven membrane system in treating sulfamethoxazole-containing waterResources Conservation and Recycling13.2哈尔滨工业大学
2024Replacing traditional pretreatment in one-step UF with natural short-distance riverbank filtration: Continuous contaminants removal and TMP increase reliefWATER RESOURCES12.8哈尔滨工业大学
2024Toward nitrogen recovery Cocultivation of microalgae and bacteria enhances the production of high value nitrogen rich cyanophycinWater Research12.8湘潭大学
2023How nutrient loading leads to alternative stable states in microbially mediated N-cycle pathways: A new insight into bioavailable nitrogen removal in urban riversWater Research12.8河海大学
2023Habitat-specific regulation of bacterial community dynamics during phytoplankton bloom succession in a subtropical eutrophic lakeWater Research12.8中国科学院
2023Dam-induced flow alternations drive the regime shift towards a cyanobacteria-dominated microbiota state in the Yangtze RiverWater Research12.8河海大学
2023Size-dependent promotion of micro(nano)plastics on the horizontal gene transfer of antibiotic resistance genes in constructed wetlandsWater Research12.8山东大学
2023Hypoxia and salinity constrain the sediment microbiota-mediated N removal potential in an estuary: A multi-trophic interrelationship perspectiveWater Research12.8河海大学
2023Phosphorus-driven regime shift from heterotrophic to autotrophic diazotrophs in a deep alpine lakeWater Research12.8中国科学院南京地理与湖泊研究所
2023Disinfectant sodium dichloroisocyanurate synergistically strengthened sludge acidogenic process and pathogens inactivation: Targeted upregulation of functional microorganisms and metabolic traits via self-adaptationWater Research12.8河海大学
2023Hydrogenotrophic pathway dominates methanogenesis along the river-estuary continuum of the Yangtze Riverwater research12.8中国科学院
2024Deforestation impacts soil biodiversity and ecosystem services
worldwide
PANS12.779南京农业大学
2023Intestinal epithelial pH-sensing receptor GPR65 maintains mucosal homeostasis via regulating antimicrobial defense and restrains gut inflammation in inflammatory bowel diseaseGut Microbes12.2同济大学医学院附属第十人民医院
2023Biodegradability enhancement of phenolic wastewater using hydrothermal pretreatmentBioresource Technology11.889安徽工业大学
2022Upgrading volatile fatty acids production from anaerobic co-fermentation of orange peel waste and sewage sludge: Critical roles of limonene on functional consortia and microbial metabolic traitsBioresource Technology11.889中国河海大学
2022Multiple hydrolyses of rice straw by domesticated paddy soil microbes for methane production via liquid anaerobic digestionBioresource Technology11.889南昌大学
2024Alternating ventilation accelerates the mineralization and humification of food waste by optimizing the temperature-oxygen-moisture distribution in the static composting reactorBioresource Technology11.4浙江大学
2024Mechanistic insights into different illumination positions control algae production in anaerobic dynamic membrane filtration (AnDM) during decentralized wastewater treatmentBioresource Technology11.4东北农业大学
2024Synergistic improvement of humus formation in compost residue by fenton-like and effective microorganism composite agentsBioresource Technology11.4四川农业大学
2023Ultrasonic-assisted hypochlorite activation accelerated volatile fatty acids production during sewage sludge fermentation: Critical insights on solubilization/hydrolysis stages and microbial traitsBioresour Technol11.4同济大学
2023Relative contribution of ammonia-oxidizing bacteria and denitrifying fungi to N2O production during rice straw composting with biochar and biogas residue amendmentsBioresour Technol11.4湖南农业大学
2023Response characteristics of antibiotic resistance genes and bacterial communities during agricultural waste composting: Focusing on biogas residue combined with biochar amendmentsBioresource Technology11.4湖南农业大学
2023Aerobic Denitrification Enhanced by Immobilized Slow-Released Iron/Activated Carbon Aquagel Treatment of Low C/N Micropolluted Water: Denitrification Performance, Denitrifying Bacterial Community Co-occurrence, and ImplicationsENVIRONMENTAL SCIENCE & TECHNOLOGY11.357西安建筑科技大学
2022Reduction of Dimethylarsenate to Highly Toxic Dimethylarsenite in Paddy Soil and Rice PlantsEnvironmental Science & Technology11.357南京农业大学
2021How environmental stress leads to alternative microbiota states in a river ecosystem: A new insight into river restorationwater Research11.239河海大学
2021Microbial community coalescence: does it matter in the Three Gorges Reservoir?water Research11.239三峡库区
2021Effects of nitrogen on the longitudinal and vertical patterns of the composition and potential function of bacterial and archaeal communities in the tidal mudflatswater Research11.239河海大学
2019Comparative analysis of the gut microbiota of the blue fox (Alopex lagopus) and raccoon dog (Nyctereutes procyonoides)water Research11.239中国动植物科学研究院
2024Distinct microbial hydrogen and reductant disposal pathways explain interbreed variations in ruminant methane yieldThe ISME Journal11.217中国科学院农业研究所
2024Methylomonadaceae was the active and dominant methanotroph in Tibet lake sedimentsISME11.217南京农业大学
2022Small changes in rhizosphere microbiome composition predict disease outcomes earlier than pathogen density variationsISME11.217南京农业大学
2022Amino acid utilization allows intestinal dominance of Lactobacillus amylovorusISME11.217南京农业大学
2022Suppression of the insect cuticular microbiomes by a fungal defensin to facilitate parasite infectionISME11.217中国科学院上海植物生理生态研究所
2023Investigation into the treatment and resource recovery of rifamycin mycelial dreg with thermal alkaline pretreatment-anaerobic digestionJournal of Cleaner Production11.1哈尔滨工业大学
2023Polyethylene microplastics can attenuate soil carbon sequestration by reducing plant photosynthetic carbon assimilation and transfer: evidence from a 13C-labeling mesocosm studyJournal of Cleaner Production11.072华南农业大学
2024Soil redox status governs within-field spatial variation in microbial arsenic methylation and rice straighthead diseaseISME J11南京农业大学
2024Effects of an efficient straw decomposition system mediated by Stropharia rugosoannulata on soil properties and microbial communities in forestlandScience of the Total Environment10.754上海市农业科学院食用菌研究所
2022Novel insights in seasonal dynamics and co-existence patterns of phytoplankton and micro-eukaryotes in drinking water reservoir, Northwest China: DNA data and ecological modelScience of the Total Environment10.753西安建筑技术大学
2022Evidence for saponin diversity–mycobiome links and conservatism of plant–fungi interaction patterns across Holarctic disjunct Panax speciesScience of The Total Environment10.753中国中医科学院&西北农林科技大学
2022Carboxylesterases from bacterial enrichment culture degrade strobilurin fungicidesScience of The Total Environment10.753北京科技大学
2022How does the polyhexamethylene guanidine interact with waste activated sludge and affect the metabolic functions in anaerobic fermentation for volatile fatty acids productionSci Total Environ10.753河海大学
2022Mix-cultured aerobic denitrifying bacteria augmented carbon and nitrogen removal for micro-polluted water: Metabolic activity, coexistence and interactions, and immobilized bacteria for reservoir raw water treatmentSci Total Environ10.753西安建筑技术大学
2022Unveiling the behaviors and mechanisms of percarbonate on the sludge anaerobic fermentation for volatile fatty acids productionSci Total Environ10.753河海大学
2022Nutrient status of integrated rice-crayfish system impacts the microbial nitrogen-transformation processes in paddy fields and rice yieldsSci Total Environ10.753河海大学
2022Co-impacts of the microplastic polyamide and sertraline on the denitrification function and microbial community structure in SBRsSci Total Environ10.753河海大学
2022Re-circulation of Fe/persulfate regulated sludge fermentation products for sewage treatment: Focus on pollutant removal efficiency, microbial community and metabolic activityScience of the Total Environment10.753安徽理工大学
2022Using heat-activated persulfate to accelerate short-chain fatty acids production from waste activated sludge fermentation triggered by sulfate-reducing microbial consortiumScience of the Total Environment10.753太原理工大学
2022Novel insights into aerobic denitrifying bacterial communities augmented denitrification capacity and mechanisms in lake watersScience of the Total Environment10.753西安建筑技术大学
2022Effects of composite materials and revegetation on soil nutrients, chemical and microbial properties in rare earth tailingsScience of the Total Environment10.753中国科学院地理科学与自然资源研究所
2020Electrochemical behavior of biochar and its effects on microbial nitrate reduc‐ tion: Role of extracellular polymeric substances in extracellular electron trans‐ ferChemical Engineering Journal10.652河海大学
2020Rhizosphere protists are key determinants of plant healthMicrobiome10.465南京农业大学
2024Effect of Streptomyces JD211 application on soil physicochemical properties and N2O emission characteristics of rice rhizosphereScience of the Total Environment9.8江西农业大学应用微生物研究所
2024The distribution profiles of tetracycline resistance genes in rice: Comparisons using four genotypesENVIRONMENTAL SCIENCES9.8扬州大学
2024The role of iron-rich organic fertilizer in promoting the growth of Chinese cabbage and inhibiting the transformation of cadmiumENVIRONMENTAL SCIENCES9.8湖南农业大学
2024Effects of an efficient straw decomposition system mediated by Stropharia
rugosoannulata on soil properties and microbial communities in forestland
Science of the Total Environment9.8上海市农业科学院
2024Deciphering the biotic and abiotic drivers of coalescence asymmetry between soil and manure microbiomesScience of the Total Environment9.8南京农业大学
2024Aggregating Synechococcus contributes to particle organic carbon export in coastal estuarine waters: Its lineage features and assembly processesScience of the Total Environment9.8中国科学院
2024Soil amendments alter cadmium distribution and bacterial community structure in paddy soilsScience of the Total Environment9.8广东省农业科学院资源与环境研究所
2024Effects of different emergent macrophytes on methane flux and
rhizosphere microbial communities in wetlands
Science of The Total Environment9.8中国科学院南京地理湖沼研究所
2024Enhanced humification of full-scale apple wood and cow manure by promoting lignocellulose degradation via biomass pretreatmentsScience of the Total Environment9.8生态环境部华南环境科学研究所
2024Environmental heterogeneity caused by large-scale cultivation of Pyropia haitanensis shapes multi-group biodiversity distribution in coastal areasScience of the Total Environment9.8集美大学水产学院
2023Phylogenetic diversity of stochasticity-dominated predatory myxobacterial community drives multi-nutrient cycling in typical farmland soilsScience of the Total Environment9.8中科院土壤科学研究所
2023Surfactant aggravated the antibiotic's stress on antibiotic resistance genes
proliferation by altering antibiotic solubilization and microbial traits in
sludge anaerobic fermentation
Science of the Total Environment9.8河海大学
2023Hydrodynamics-driven community coalescence determines ecological assembly processes and shifts bacterial network stability in river bendsScience of the Total Environment9.8河海大学
2023Promoted aerobic denitrification through denitrifying fungal communities: Co-occurrence patterns and treatment of low C/N micro-polluted waterScience of the Total Environment9.8西安建筑科技大学
2023Surfactant and antibiotic co-occurrence reshaped the acidogenic process for
volatile fatty acids production during sludge anaerobic fermentation
Science of the Total Environment9.8河海大学
2023Mitigating the adverse effect of warming on rice canopy and rhizosphere microbial community by nitrogen application: An approach to counteract future climate change for riceScience of the Total Environment9.8南京农业大学
2023Distinct effects of chemical- and bio- flocculants on the sludge acidogenic fermentation for volatile fatty acids production by affecting the acidogenic steps, microbial community structure and metabolic functionsScience of the Total Environment9.8同济大学
2023Acid rain reduces plant-photosynthesized carbon sequestration and soil microbial network complexitySCIENCE OF THE TOTAL ENVIRONMENT9.8华南农业大学
2023Nutrient strengthening of winter wheat by foliar ZnO and Fe3O4 NPs: Food safety, quality, elemental distribution and effects on soil bacteriaScience of the Total Environment9.8西北农林科技大学
2022Mix-Cultured Aerobic Denitrifying Bacterial Communities Augmented Treatment of Micro-Polluted Landscape Water: Performance, Mechanism, and ApplicationsSci Total Environ9.8西安建筑技术大学
2024Evidence of complete ammonia-oxidizing microbial communities and their contribution to N2O emissions in typical vegetable fields across ChinaSoil Biology and Biochemistry9.7南京农业大学
2023Alterations in substrate stoichiometry control the responses of soil diazotrophs to nutrient enrichmentSoil Biology and Biochemistry9.7南京农业大学
2022Nitrogen removal from low carbon/nitrogen polluted water is enhanced by a novel synthetic micro-ecosystem under aerobic conditions: Novel insight into abundance of denitrification genes and community interactionsBioresource Technology9.642西安建筑科技大学
2021Ensiling pretreatment with two novel microbial consortia enhances bioethanol production in sterile rice strawBioresour Technol9.641南京农业大学
2021Use of biochar to reduce mercury accumulation in Oryza sativa L: A trial for sustainable management of historically polluted farmlandsEnvironment International9.6中国科学院地球化学研究所
2023Microbiome and spatially resolved metabolomics analysis reveal the anticancer
role of gut Akkermansia muciniphila by crosstalk with intratumoral microbiota
and reprogramming tumoral metabolism in mice
GUT MICROBES9.434同济大学医学院
2019Butyric acid normalizes hyperglycemia caused by the tacrolimus-induced gut microbiotaAm J Transplant9.369首都医科大学附属医院
2022Function of aquatic plants on nitrogen removal and greenhouse gas emission in enhanced denitrification constructed wetlands: Iris pseudacorus for exampleJournal of Cleaner Production9.297上海交通大学
2022Reutilization of waste crawfish shell and sludge for efficient volatile fatty acids production by synchronously regulating the bioavailable substrates and microbial metabolic traitsJournal of Cleaner Production9.297河海大学
2022Polyethylene microplastics can attenuate soil carbon sequestration by reducing plant photosynthetic carbon assimilation and transfer: evidence from a 13C-labeling mesocosm studyJournal of Cleaner Production9.297华南农业大学
2021Erosion reduces soil microbial diversity, network complexity and multifunctionalityISME9.181西北农林科技大学
2019organophosphorus pesticide chlorpyrifos intake promotes obesity and insulin resisitance through impacting gut and gut microbiotaMicrobiome9.133中国农业大学
2019Antibiotics may increase triazine herbicide exposure risk via disturbing gut microbiotaMicrobiome9.133中国农业大学
2019Ruminal microbiome-host crosstalk stimulates the development of the ruminal epithelium in a lamb modelMicrobiome9.133南京农业大学
2019Competitive interaction with keystone taxa induced negative priming under biochar amendmentsMicrobiome9.133中国科学院土壤研究所
2020New insights into the vertical distribution and microbial degradation of microplastics in urban river sedimentswater research9.13河海大学
2020Dams shift microbial community assembly and imprint nitrogen transformation along the Yangtze Riverwater research9.13河海大学
2020Grain size tunes microbial community assembly and nitrogen transformation activity under frequent hyporheic exchange: A column experimentwater Research9.13河海大学
2022Production of Helvolic Acid in Metarhizium Contributes to Fungal Infection of Insects by Bacteriostatic Inhibition of the Host Cuticular MicrobiomesMicrobiology Spectrum9.043中国科学院上海植物生理生态研究所


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上海凌恩生物科技有限公司(简称“凌恩生物”)成立于2014年底,是一家从事生物科学研究技术服务的高科技公司,专注于二代高通量测序及三代单分子测序技术在科研、农业发展及人类健康领域的研究和应用。

公司成立至今,凭借IlluminaPacBio公司的二代及三代测序平台开展基因组学测序及生信分析服务,销售网络遍布全国,甚至为美国、德国、英国、日本、新加坡、马来西亚、也门等地的国际客户实现了跨国服务。秉持“以客户需求为本、为客户创造价值”的服务宗旨,以高品质、高效率的特色技术服务,获得科研用户的广泛赞誉,合作伙伴遍及全球。


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