Metagenomic Sequencing-Nanopore
Kev Pabcuam Zoo
● Kev sib dhos zoo-Txhim kho qhov tseeb ntawm hom kev txheeb xyuas thiab kev ua haujlwm ntawm cov noob kwv yees
● Kaw cov kab mob genome cais
● Cov ntawv thov muaj zog dua thiab txhim khu kev qha hauv ntau thaj chaw, xws li kuaj pom cov kab mob pathogenic lossis cov tshuaj tiv thaiv kab mob ntsig txog cov noob.
● Sib piv metagenome tsom xam
Service Specifications
| Platform | Sequencing | Cov ntaub ntawv pom zoo | Lub sijhawm hloov pauv |
| Nanopor | ONT | 6 G / 10 G | 65 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 thiab kev xa khoom
Cov Qauv Xav Tau:
RauDNA rho tawm:
| Hom qauv | Tus nqi | Nco ntsoov | Purity |
| DNA rho tawm | Yog '- 1.5 g | 20 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
1.Heatmap: Hom kev nplua nuj pawg
2.Cov noob ua haujlwm tau sau tseg rau KEGG metabolic txoj hauv kev
3.Species correlation network
4.Circos ntawm CARD tshuaj tiv thaiv kab mob
BMK Cas
Nanopore metagenomics ua rau kev kuaj mob sai sai ntawm cov kab mob hauv qab ua pa
Luam tawm:Ntuj Biotechnology, 2019
Technical Highlights
Sequencing: Nanopore MinION
Clinical metagenomics bioinformatics: Tus tswv tsev DNA depletion, WIMP thiab ARMA tsom xam
Kev kuaj pom sai: 6 teev
Siab rhiab heev: 96.6%
Cov txiaj ntsig tseem ceeb
Xyoo 2006, tus kab mob ua pa qis (LR) ua rau 3 lab tib neeg tuag thoob ntiaj teb.Txoj kev zoo tshaj plaws rau LR1 kev kuaj kab mob yog kev cog qoob loo, uas muaj qhov tsis zoo, lub sijhawm tig mus ntev thiab tsis muaj kev taw qhia hauv kev kho tshuaj tua kab mob thaum ntxov.Kev kuaj mob sai thiab raug microbial tau ntev lawm yog qhov xav tau ceev.Dr. Justin los ntawm University of East Anglia thiab nws cov neeg koom tes ua tiav tau tsim ib txoj kev Nanopore-based metagenomic rau kev kuaj kab mob.Raws li lawv cov haujlwm ua haujlwm, 99.99% ntawm tus tswv DNA tuaj yeem ua tiav.Kev kuaj xyuas cov kab mob thiab cov tshuaj tiv thaiv kab mob tuaj yeem ua tiav hauv 6 teev.
Kev siv
Charalampous, T., Kay, GL, Richardson, H., Aydin, A., & O'Grady, J. .(2019).Nanopore metagenomics ua rau kev kuaj mob sai sai ntawm cov kab mob hauv qab ua pa.Ntuj Biotechnology, 37(7), 1.
