16S/18S/ITS Amplicon Sequencing-NGS
Amfanin Sabis
● Warewa-kyauta da saurin gano abubuwan ƙwayoyin cuta a cikin samfuran muhalli
● Babban ƙuduri a cikin ƙananan abubuwa masu yawa a cikin samfuran muhalli
● Sabbin QIIME2 na nazarin kwarara tare da nazari iri-iri dangane da bayanan bayanai, annotation, OTU/ASV.
● Babban kayan aiki, daidaito mafi girma
● Ana amfani da su don nazarin ƙananan ƙwayoyin cuta daban-daban
● BMK mallaki m kwarewa tare da kan 100,000 samfurori / shekara ， rufe ƙasa, ruwa, gas, sludge, feces, hanji, fata, fermentation broth, kwari, shuke-shuke, da dai sauransu.
BMKCloud ya sauƙaƙe fassarar bayanai mai ɗauke da kayan aikin tantance keɓaɓɓen mutum 45
Ƙayyadaddun Sabis
| JeriDandalin | Laburare | Nasihar yawan amfanin bayanai | Kiyasin lokacin juyawa |
| Illumina NovaSeq Platform | PE250 | 50K/100K/300K Tags | Kwanaki 30 |
Binciken bioinformatics
● Raw data ingancin kula
● OTU clustering/De-noise(ASV)
● Bayanin OTU
● Bambancin Alpha
● Bambancin Beta
● Bincike tsakanin ƙungiyoyi
● Binciken ƙungiyar akan abubuwan gwaji
● Hasashen kwayoyin aiki
Samfuran Bukatun da Bayarwa
Samfuran Bukatun:
DominCire DNA:
| Nau'in Misali | Adadin | Hankali | Tsafta |
| Cire DNA | ku 30ng | 1 ng/l | OD260/280= 1.6-2.5 |
Don samfuran muhalli:
| Nau'in samfurin | Hanyar samfurin da aka ba da shawarar |
| Ƙasa | Adadin samfur: kimanin.5 g;ku.Ya kamata a cire ragowar abin da ya bushe daga saman;Niƙa manyan guda kuma ku wuce ta 2 mm tace;Samfuran Aliquot a cikin bakararre EP-tube ko cyrotube don ajiya. |
| Najasa | Adadin samfur: kimanin.5 g;ku.Tattara da liquot samfurori a cikin bakararre EP-tube ko cryotube don ajiyar wuri. |
| Abubuwan ciki na hanji | Ana buƙatar sarrafa samfuran ƙarƙashin yanayin aseptic.Wanke nama da aka tattara tare da PBS;Centrifuge PBS kuma tattara hazo a cikin EP-tuben. |
| Lalacewa | Adadin samfur: kimanin.5 g;ku.Tattara da liquot samfurin sludge a cikin bakararre EP-tube ko cryotube don ajiya |
| Ruwan ruwa | Don samfurin tare da iyakataccen adadin ƙwayoyin cuta, kamar ruwan famfo, ruwan rijiya, da sauransu, Tattara ruwa aƙalla 1 L kuma ku wuce ta 0.22 μm tace don wadatar da ƙananan ƙwayoyin cuta akan membrane.Ajiye membrane a cikin bututu bakararre. |
| Fatar jiki | A hankali a goge saman fata tare da swab ɗin auduga mai bakararre ko ɓawon tiyata sannan a sanya shi cikin bututu mara kyau. |
Isar da Samfurin Nasiha
Daskare samfuran a cikin ruwa nitrogen na sa'o'i 3-4 kuma adana a cikin ruwa nitrogen ko -80 digiri zuwa ajiyar dogon lokaci.Ana buƙatar jigilar samfuri tare da bushe-kankara.
Gudun Aikin Sabis
Samfurin bayarwa
Gina ɗakin karatu
Jeri
Binciken bayanai
Bayan-sayar da sabis
1.Species rarraba
2.Zafi taswira: Nau'in wadata tari
3.Rare bangaran lankwasa
4.NMDS bincike
5.Lefse bincike
BMK Case
Mutanen da ke da nau'in ciwon sukari na 2 da ba tare da su ba suna nuna ƙarfin aikin ƙwayoyin cuta daban-daban da abun da ke ciki.
Buga:Mai watsa shiri na Cell & Microbe, 2019
Dabarun tsarawa:
Lean mara ciwon sukari (n=633);Marasa kiba mai kiba (n=494);Kiba-Nau'in ciwon sukari na 2 (n=153);
Yankin manufa: 16S rDNA V1-V2
Platform: Illumina Miseq (tsarin amplicon na tushen NGS)
An ƙaddamar da ɓangarori na tsantsa DNA zuwa jeri na metagenomic akan Illumina Hiseq
Sakamako mai mahimmanci
An yi nasarar bambanta bayanan ƙananan ƙwayoyin cuta na waɗannan cututtuka na rayuwa.
Ta hanyar kwatanta sifofin ƙananan ƙwayoyin cuta waɗanda aka samar ta hanyar jerin 16S, an gano kiba don haɗuwa da canje-canje a cikin abubuwan microbial, siffofi na mutum, musamman raguwa a Akkermansia, Faecalibacterium, Oscillibacter, Alistipes, da dai sauransu. Bugu da ƙari, an samo T2D tare da karuwa a cikin Escherichia / shigella. .
Magana
Thingholm, LB, et al."Mutane masu Kiba tare da Ba tare da Ciwon Ciwon sukari Na 2 ba suna Nuna Ƙarfin Ayyukan Kwayoyin Kwayoyin cuta daban-daban."Mai watsa shiri na Cell & Microbe26.2 (2019).
