Metagenomic Sequencing -NGS

Metagenome yog hais txog kev sau tag nrho cov khoom siv caj ces ntawm cov zej zog sib xyaw ntawm cov kab mob, xws li ib puag ncig metagenome, tib neeg metagenome, thiab lwm yam. Nws muaj cov genomes ntawm ob qho tib si cultivatable thiab uncultivatable microorganisms.Metagenomic sequencing yog cov cuab yeej molecular siv los txheeb xyuas cov khoom sib xyaw genomic muab rho tawm los ntawm ib puag ncig cov qauv, uas muab cov ncauj lus kom ntxaws txog hom kev sib txawv thiab kev nplua nuj, cov qauv pej xeem, kev sib raug zoo ntawm phylogenetic, cov noob caj noob ces thiab kev sib raug zoo nrog ib puag ncig.

Platform:Illumina NovaSeq 6000

Tiv tauj peb

Kev Pabcuam Paub meej

Cov txiaj ntsig Demo

Case Study

Kev Pabcuam Zoo

● Kev rho tawm thiab cog qoob loo-dawb rau microbial zej zog profileing

● Kev daws teeb meem siab hauv kev txheeb xyuas cov tsiaj tsis tshua muaj nyob hauv cov qauv ib puag ncig

● Lub tswv yim ntawm "meta-" integrates tag nrho cov yam ntxwv lom ntawm kev ua haujlwm, qib hom thiab qib noob, uas qhia txog kev pom zoo uas ze rau qhov tseeb.

● BMK sau cov kev paub loj hauv ntau hom qauv nrog ntau dua 10,000 cov qauv ua tiav.

Service Specifications

Platform	Sequencing	Cov ntaub ntawv pom zoo	Lub sijhawm hloov pauv
Illumina NovaSeq 6000 Nws	PEB 150	6 G / 10 G / 20 G	45 hnub ua haujlwm

Bioinformatics tsom xam

● Raw cov ntaub ntawv zoo tswj

● Metagenom los ua ke

● Cov noob caj noob ces thiab cov lus piav qhia uas tsis yog redundant

● Kev txheeb xyuas qhov sib txawv

● Genetic function diversity tsom xam

● Kev tsom xam ntawm pab pawg

● Kev txheeb xyuas kev sib koom ua ke tawm tsam qhov kev sim

Sample Requirements thiab Delivery

Cov Qauv Xav Tau:

RauDNA rho tawm:

Hom qauv	Tus nqi	Nco ntsoov	Purity
DNA rho tawm	＞ 100 ng	1 ng / μl	OD 260/280 = 1.6-2.5

Rau ib puag ncig cov qauv:

Hom qauv	Cov txheej txheem kuaj pom pom zoo
Av	Sampling tus nqi: approx.5 g yog ';Cov khoom uas tseem tshuav yuav tsum tau muab tshem tawm ntawm qhov chaw;Sib tsoo cov khoom loj thiab hla dhau 2 hli lim;Aliquot kuaj hauv sterile EP-tube lossis cyrotube rau kev tshwj tseg.
Cov quav	Sampling tus nqi: approx.5 g yog ';Sau thiab aliquot cov qauv hauv sterile EP-tube lossis cryotube rau kev tshwj tseg.
Cov ntsiab lus plab hnyuv	Cov qauv yuav tsum tau ua tiav nyob rau hauv aseptic mob.Ntxuav cov ntaub so ntswg nrog PBS;Centrifuge PBS thiab sau cov dej nag hauv EP-tubes.
Sludge	Sampling tus nqi: approx.5 g yog ';Sau thiab aliquot sludge qauv hauv sterile EP-tube lossis cryotube rau kev tshwj tseg
Tus dej	Rau cov qauv uas txwv tsis pub muaj microbial, xws li dej kais dej, dej zoo, thiab lwm yam, Sau tsawg kawg yog 1 L dej thiab dhau los ntawm 0.22 μm lim kom txhawb microbial ntawm daim nyias nyias.Khaws cov membrane rau hauv lub raj tsis muaj menyuam.
Daim tawv nqaij	Ua tib zoo txhuam cov tawv nqaij nrog cov paj rwb sterile swab los yog phais hniav thiab muab tso rau hauv sterile raj.

Pom zoo coj mus kuaj

Kho cov qauv hauv cov kua nitrogen rau 3-4 teev thiab khaws cia hauv cov kua nitrogen los yog -80 degree mus rau lub sijhawm ntev.Qauv shipping nrog qhuav-dej khov yog yuav tsum tau.

Kev Pabcuam Ua Haujlwm Flow

Qauv xa tuaj

Kev tsim kho tsev qiv ntawv

Sequencing

Cov ntaub ntawv tsom xam

Kev pab cuam tom qab muag

Yav dhau los: Metagenomic Sequencing-Nanopore

Tom ntej: Chromatin Immunoprecipitation Sequencing (ChIP-seq)

1.Histogram: Hom kev faib tawm

2.Cov noob ua haujlwm tau sau tseg rau KEGG metabolic txoj hauv kev

3.Heat map: Differential functions raws li cov txheeb ze cov noob abundance4.Circos ntawm CARD tshuaj tiv thaiv kab mob

BMK Cas

Feem ntau ntawm cov tshuaj tiv thaiv kab mob thiab cov kab mob pathogens raws cov av-mangrove cag continuum

Luam tawm:Phau ntawv Journal of Hazardous Materials, 2021

Sequence Strategy:

Cov ntaub ntawv: DNA rho tawm ntawm plaub feem ntawm mangrove cag txuam cov qauv: unplanted av, rhizosphere, episphere thiab endosphere compartments
Platform: Illumina HiSeq 2500
Hom phiaj: Metagenome
16S rRNA gene V3-V4 cheeb tsam

Cov txiaj ntsig tseem ceeb

Metagenomic sequencing thiab metabarcoding profiling ntawm cov av hauv paus txuas ntxiv ntawm mangrove saplings tau ua tiav txhawm rau kawm txog kev nthuav tawm cov tshuaj tiv thaiv kab mob (ARGs) los ntawm cov av rau hauv cov nroj tsuag.Metagenomic cov ntaub ntawv qhia tau hais tias 91.4% ntawm cov tshuaj tiv thaiv kab mob yog feem ntau pom nyob rau hauv tag nrho plaub cov av compartments hais saum toj no, uas qhia ib tug tas mus li zam.16S rRNA amplicon sequencing generated 29,285 sequences, sawv cev rau 346 hom.Ua ke nrog cov tsiaj ntawv profileing los ntawm amplicon sequencing, qhov kev nthuav tawm no tau pom tias muaj kev ywj pheej ntawm cov hauv paus-koom nrog microbiota, txawm li cas los xij, nws tuaj yeem txhawb nqa los ntawm cov xov tooj ntawm cov caj ces.Txoj kev tshawb no tau txheeb xyuas qhov ntws ntawm ARGs thiab cov kab mob los ntawm cov av mus rau hauv cov nroj tsuag los ntawm kev sib txuas hauv av-hauv paus txuas ntxiv.

Kev siv

Wang, C., Hu, R., Strong, PJ, Zhuang, W., & Shu, L. .(2020).Feem ntau ntawm cov tshuaj tiv thaiv kab mob thiab cov kab mob pathogens raws cov av-mangrove cag continuum.Phau ntawv Journal of Hazardous Materials, 408Ib., 124985.