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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。

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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(两大技术平台)

Illunima测序:

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PacBio测序(扩增子全长测序):

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内容

项目引物名称序列扩增产物长度
全长

16S 全长

27F5′-AGRGTTTGATYNTGGCTCAG-3′1.5K
1492R5′-TASGGHTACCTTGTTASGACTT-3′1.5K

18S全长

EukA5′-AACCTGGTTGATCCTGCCAGT-3′1.8K
EukB5′-GATCCTTCTGCAGGTTCACCTAC-3′1.8K

ITS 全长

ITS15′- CTTGGTCATTTAGAGGAAGTAA-3′0.6-0.7K
ITS45′- TCCTCCGCTTATTGATATGC-3′0.6-0.7K



分析流程

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NGS扩增子测序

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PacBio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≥150ng

2)浓度≥5ng/ul



技术特色

  • 两大测序平台,满足不同多样性检测需求

  • 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


二代多样性

发表时间

文章标题杂志名影响因子合作单位

2023

Biodegradability enhancement of phenolic wastewater using hydrothermal pretreatment

Bioresource Technology

11.889

安徽工业大学

2023

Polyethylene microplastics can attenuate soil carbon sequestration by reducing plant photosynthetic carbon assimilation and transfer: evidence from a 13C-labeling mesocosm study

Journal of Cleaner Production

11.072

华南农业大学

2023

Phylogenetic diversity of stochasticity-dominated predatory myxobacterial community drives multi-nutrient cycling in typical farmland soils

Science of the Total Environment

10.753

中国科学院

2023

Hydrodynamics-driven community coalescence determines ecological assembly processes and shifts bacterial network stability in river bends

Science of the Total Environment

10.753

河海大学

2022

Antimicrobial PCMX facilitates the volatile fatty acids production during sludge anaerobic fermentation: Insights of the interactive principles, microbial metabolic profiles and adaptation mechanisms

Chemical Engineering Journal

16.744

河海大学

2022

Enhanced removal of organic, nutrients, and PFCs in the iron-carbon micro-electrolysis constructed wetlands: mechanism and iron cycle

Chemical Engineering Journal

16.744

河海大学

2022

Soil particle size fractions affect arsenic (As) release and speciation: insights into dissolved organic matter and functional genes

Journal of Hazardous Materials

14.224

华南环境科学研究所

2022

Diversity, function and assembly of the Trifolium repens L. root-associated microbiome under lead stress

Journal of Hazardous Materials

14.224

东北农业大学

2022

Toward emerging contaminants removal using acclimated activated sludge in the gravity-driven membrane filtration system

Journal of Hazardous Materials

14.224

哈尔滨工业大学

2022

Insight into the role of biogenic manganese oxides-assisted gravity-driven membrane filtration systems toward emerging contaminants removal

Water Research

13.4

哈尔滨工业大学

2022

Sustainable restoration of anoxic freshwater using environmentally-compatible oxygen-carrying biochar: Performance and mechanisms

Water Research

13.4

河海大学

2022

Di-(2-ethylhexyl) phthalate exacerbated the toxicity of polystyrene nanoplastics through histological damage and intestinal microbiota dysbiosis in freshwater Micropterus salmoides

Water Research

13.4

华南农业大学

2022

Combined remediation effects of biochar and organic fertilizer on immobilization and dissipation of neonicotinoids in soils

Environment International

13.352

扬州大学

2022

Upgrading 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 traits

Bioresource Technology

11.889

中国河海大学

2022

Multiple hydrolyses of rice straw by domesticated paddy soil microbes for methane production via liquid anaerobic digestion

Bioresource Technology

11.889

南昌大学

2022

Reduction of Dimethylarsenate to Highly Toxic Dimethylarsenite in Paddy Soil and Rice Plants

Environmental Science & Technology

11.357

南京农业大学

2022

Small changes in rhizosphere microbiome composition predict disease outcomes earlier than pathogen density variations

ISME

11.217

南京农业大学

2022

Amino acid utilization allows intestinal dominance of Lactobacillus amylovorus

ISME

11.217

南京农业大学

2022

Suppression of the insect cuticular microbiomes by a fungal defensin to facilitate parasite infection

ISME

11.217

中国科学院

2022

Evidence for saponin diversity–mycobiome links and conservatism of plant–fungi interaction patterns across Holarctic disjunct Panax species

Science of The Total Environment

10.753

中国中医科学院,

西北农林科技大学

2022

How does the polyhexamethylene guanidine interact with waste activated sludge and affect the metabolic functions in anaerobic fermentation for volatile fatty acids production

Sci Total Environ

10.753

河海大学

2022

Unveiling the behaviors and mechanisms of percarbonate on the sludge anaerobic fermentation for volatile fatty acids production

Sci Total Environ

10.753

河海大学

2022

Nutrient status of integrated rice-crayfish system impacts the microbial nitrogen-transformation processes in paddy fields and rice yields

Sci Total Environ

10.753

河海大学

2022

Co-impacts of the microplastic polyamide and sertraline on the denitrification function and microbial community structure in SBRs

Sci Total Environ

10.753

河海大学

2022

Re-circulation of Fe/persulfate regulated sludge fermentation products for sewage treatment: Focus on pollutant removal efficiency, microbial community and metabolic activity

Science of the Total Environment

10.753

安徽理工大学

2022

Using heat-activated persulfate to accelerate short-chain fatty acids production from waste activated sludge fermentation triggered by sulfate-reducing microbial consortium

Science of the Total Environment

10.753

太原理工大学

2022

Novel insights into aerobic denitrifying bacterial communities augmented denitrification capacity and mechanisms in lake waters

Science of the Total Environment

10.753

西安建筑技术大学

2022

Effects of composite materials and revegetation on soil nutrients, chemical and microbial properties in rare earth tailings

Science of the Total Environment

10.753

中国科学院

2021

How environmental stress leads to alternative microbiota states in a river ecosystem: A new insight into river restoration

water Research

11.239

河海大学

2021

Microbial community coalescence: does it matter in the Three Gorges Reservoir?

water Research

11.239

三峡库区

2021

Effects of nitrogen on the longitudinal and vertical patterns of the composition and potential function of bacterial and archaeal communities in the tidal mudflats

water Research

11.239

河海大学

三代多样性

2023

Short-term response of soil enzyme activities and bacterial communities in black soil to a herbicide mixture: Atrazine and Acetochlor

Applied Soil Ecology

5.509

东北农业大学

2023

Polymeric immunoglobulin receptor deficiency exacerbates autoimmune hepatitis by inducing intestinal dysbiosis and barrier dysfunction

Cell Death and Disease

9.685

温州医科大学第一附属医院

2023

Monitoring and identifi cation of spoilage-related microorganisms in braised chicken with modifi ed atmosphere packaging during refrigerated storage

Food Science and Human Wellness

8.022

南京农业大学

2023

Anti-virulence activity of novel (1-heteroaryloxy-2- hydroxypropyl)-   phenylpiperazine derivatives against both wild-type and clinical drug-resistant Candida albicansstrains

Microbial Biotechnology

6.575

广州医科大学

2023

Clostridium butyricum Prevents Dysbiosis and the Rise in Blood Pressure in Spontaneously Hypertensive Rats

International Journal of Molecular Sciences

5.923

中国海洋大学

2023

Traditional Chinese medicines (TCMs) with varied meridians (Gui-Jing) differentially alleviate the adverse impact of Coptis chinensis on gut microbiota

Journal of Ethnopharmacology

5.195

天津中医药大学中医学院

2022

ETS-1 facilitates Th1 cell-mediated mucosal inflammation in inflammatory bowel diseases through upregulating CIRBP

Journal of Autoimmunity

14.511

同济大学医学院上海第十人民医院

2022

Bacterial dynamics of drinking water pipeline: A new perspective on dual effects of stagnation and algal organic matter

Water Research

13.4

西安建筑技术大学

2022

Bacterial community structure and metabolic activity of drinking water pipelines in buildings: A new perspective on dual effects of hydrodynamic stagnation and algal organic matter invasion

Water Research

13.4

西安建筑技术大学

2022

Small Intestine-specific Knockout of CIDEC Improves Obesity and Hepatic Steatosis by Inhibiting Synthesis of Phosphatidic Acid

International Journal of Molecular Sciences

10.75

广西大学

2022

Linarin ameliorates dextran sulfate sodium-induced colitis in C57BL/6J mice via the improvement of intestinal barrier, suppression of inflammatory responses and modulation of gut microbiota

Food & Function

6.317

西北农林科技大学

2022

Heat Stress Weakens the Skin Barrier Function in Sturgeon by Decreasing Mucus Secretion and Disrupting the Mucosal Microbiota.

Frontiers in Microbiology

6.064

四川农业大学

2022

The Gut Microbial Co-Abundance Gene Groups (CAGs) Differentially Respond to the Flavor (Yao-Wei) of Chinese Meteria Medica

American Journal of Chinese Medicine

6.005

天津中医药大学

电 话:021-31021022

            13032137192

邮 箱:lab@biozeron.com


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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