Tsarin Metagenomic -NGS
Amfanin Sabis
● Keɓancewa da noma-kyau don ƙayyadaddun bayanan ƙananan ƙwayoyin cuta
● Babban ƙuduri a cikin gano ƙananan nau'in nau'in nau'in nau'in nau'in nau'i a cikin samfurori na muhalli
● Ma'anar "meta-" ta haɗa dukkan siffofi na halitta a matakin aiki, matakin jinsin da kuma matakin jinsin halitta, wanda ke nuna ra'ayi mai ƙarfi wanda ya fi kusa da gaskiya.
● BMK yana tara ƙwarewa mai yawa a cikin nau'ikan samfuri daban-daban tare da samfuran sama da 10,000 da aka sarrafa.
Ƙayyadaddun Sabis
| Dandalin | Jeri | Bayanan da aka ba da shawarar | Lokacin juyawa |
| Illumina NovaSeq Platform | PE150 | 6 G/10 G/20 G | 45 Kwanakin aiki |
Binciken bioinformatics
● Raw data ingancin kula
● Metagenome taro
● Saitin kwayoyin halitta da ba a sake sakewa ba
● Nazari iri-iri
● Binciken bambancin aikin kwayoyin halitta
● Bincike tsakanin ƙungiyoyi
● Binciken ƙungiyar 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;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.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 Materials masu haɗari, 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 mai mahimmanci
Matsakaicin metagenomic da bayanan metabarcoding akan tushen ƙasa na ci gaba da tsiron mangrove an sarrafa su don yin nazarin yaduwar ƙwayoyin ƙwayoyin cuta na rigakafi (ARGs) daga ƙasa zuwa tsirrai.Bayanai na metagenomic sun nuna cewa kashi 91.4% na kwayoyin juriya na ƙwayoyin cuta an gano su a cikin dukkan sassan ƙasa huɗu da aka ambata a sama, waɗanda ke nuna yanayin ci gaba.16S rRNA amplicon sequencing ya haifar da jeri 29,285, wakiltar nau'ikan 346.Haɗuwa da nau'ikan bayanin nau'ikan ta hanyar tsarin amplicon, an gano wannan watsawar ta kasance mai zaman kanta daga tushen microbiota mai alaƙa, duk da haka, ana iya sauƙaƙe ta ta wayar hannu na abubuwan 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 ƙasa-mangrove tushen ci gaba.Jaridar Materials masu haɗari, 408, 124985.
