Tsarin Metagenomic (NGS)
Amfanin Sabis
ØWarewa da noma-kyauta don bayyanar da ƙananan ƙwayoyin cuta
ØBabban ƙuduri a gano ƙananan nau'in nau'in nau'in yawa a cikin samfuran muhalli
ØMa'anar "meta-" ta haɗa dukkan siffofi na halitta a matakin aiki, nau'in nau'in jinsin da jinsin halitta, wanda ke nuna ra'ayi mai mahimmanci wanda ya fi kusa da gaskiya.
ØBMK yana tara ƙwarewa mai yawa a cikin nau'ikan samfuri daban-daban tare da samfuran sama da 10,000 da aka sarrafa.
Ƙayyadaddun Sabis
| JeriDandalin | Laburare | Nasihar yawan amfanin bayanai | Kiyasin lokacin juyawa |
| Illumina NovaSeq 6000 | PE250 | 50K/100K/300K Tags | Kwanaki 30 |
Binciken bioinformatics
üRaw data ingancin iko
üMetagenome taro
üSaitin kwayoyin halitta marasa ƙarfi da annotation
üNazari iri-iri
üBinciken bambancin aikin kwayoyin halitta
üBinciken Inter-group
üBinciken ƙungiyoyi akan abubuwan gwaji
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;Aliquot samfurori 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-tubes. |
| Lalacewa | Adadin samfur: kimanin.5 g; ku.Tattara da liquot samfurin sludge a cikin bakararre EP-tube ko cryotube don ajiyar wuri |
| 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 bakararre auduga ko ruwan fida da sanya shi cikin bututu maras 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.Histogram: Rarraba nau'ikan
2.Ayyukan kwayoyin halitta annotate zuwa KEGG hanyoyin rayuwa
3.Heat taswira: Ayyuka daban-daban dangane da yawan jinsin dangi
4.Circos na CARD kwayoyin juriya
BMK Case
Yawaitar kwayoyin juriya na ƙwayoyin cuta da ƙwayoyin cuta tare da ci gaba da tushen ƙasa-mangrove
Buga:Jaridar Abubuwan Matsala, 2021
Dabarun tsarawa:
Materials: DNA tsantsa guda hudu na tushen mangrove samfurori masu alaƙa: ƙasa marar shuka, rhizosphere, episphere da sassan endosphere
Platform: Illumina HiSeq 2500
Manufa: Metagenome
16S rRNA gene V3-V4 yankin
Sakamako masu mahimmanci
Matsakaicin metagenomic da bayanan metabarcoding akan tushen ƙasa na ci gaba da tsiron mangrove an sarrafa su don yin nazarin yaduwar ƙwayoyin ƙwayoyin ƙwayoyin cuta (ARGs) daga ƙasa zuwa tsirrai.Bayanan metagenomic sun nuna cewa kashi 91.4% na kwayoyin juriya na ƙwayoyin cuta yawanci ana gano su a duk sassan ƙasa huɗu da aka ambata a sama, waɗanda ke nuna salon ci gaba.16S rRNA amplicon sequencing ya haifar da jeri 29,285, wakiltar nau'ikan 346.Haɗuwa da nau'in sifofi na nau'in ta hanyar amplicon sequencing, an gano wannan watsawar ta kasance mai zaman kanta daga tushen microbiota mai alaƙa, duk da haka, ana iya sauƙaƙe ta hanyar wayar salula na kwayoyin halitta.Wannan binciken ya gano kwararar ARGs da ƙwayoyin cuta daga ƙasa zuwa cikin tsire-tsire ta hanyar ci gaba da tushen ƙasa mai alaƙa.
Magana
Wang, C., Hu, R., Mai ƙarfi, PJ, Zhuang, W., & Shu, L..(2020).Yawaitar kwayoyin juriya na ƙwayoyin cuta da ƙwayoyin cuta tare da tushen ƙasa-mangrove ci gaba.Jaridar Materials masu haɗari, 408, 124985.
