ØKudzipatula komanso kulima kopanda mbiri yamagulu ang'onoang'ono
ØKusamvana kwakukulu pakuzindikira mitundu yocheperako mu zitsanzo zachilengedwe
ØLingaliro la "meta-" limagwirizanitsa zinthu zonse zamoyo pamlingo wogwirira ntchito, mlingo wa mitundu ndi jini, zomwe zimasonyeza malingaliro amphamvu omwe ali pafupi ndi zenizeni.
ØBMK imadziunjikira zambiri mumitundu yosiyanasiyana yokhala ndi zitsanzo zopitilira 10,000 zokonzedwa.
Kutsatansanja | Library | Zokolola zovomerezeka | Nthawi yoyerekeza yozungulira |
Illumina NovaSeq 6000 | Chithunzi cha PE250 | 50K/100K/300K Tags | Masiku 30 |
üKuwongolera khalidwe la data yaiwisi
üMsonkhano wa Metagenome
üMa jini osagwirizana ndi ma jini ndi ndemanga
üKusanthula kwamitundu yosiyanasiyana
üKusanthula kwamitundu yosiyanasiyana ya ma genetic
üKusanthula kwamagulu
üKusanthula kwamagulu motsutsana ndi zinthu zoyeserera
ZaZithunzi za DNA:
Mtundu Wachitsanzo | Ndalama | Kukhazikika | Chiyero |
Zithunzi za DNA | > 30 ng | > 1 ng/μl | OD260/280= 1.6-2.5 |
Za zitsanzo zachilengedwe:
Mtundu wachitsanzo | Njira yoyeserera yolangizidwa |
Nthaka | Kuchuluka kwa zitsanzo: pafupifupi.5 g pa;Chotsalira chofota chiyenera kuchotsedwa pamwamba;Pewani zidutswa zazikulu ndikudutsa fyuluta ya 2 mm;Zitsanzo za Aliquot mu chubu chosabala cha EP kapena cyrotube kuti zisungidwe. |
Ndowe | Kuchuluka kwa zitsanzo: pafupifupi.5 g pa;Sonkhanitsani ndi aliquot zitsanzo mu EP-chubu wosabala kapena cryotube kuti musungidwe. |
Zomwe zili m'matumbo | Zitsanzo ziyenera kukonzedwa pansi pa chikhalidwe cha aseptic.Sambani minofu yosonkhanitsidwa ndi PBS;Centrifuge the PBS ndi kusonkhanitsa precipitant mu EP-machubu. |
Sludge | Kuchuluka kwa zitsanzo: pafupifupi.5 g pa;Sungani ndi aliquot sludge zitsanzo mu wosabala EP-chubu kapena cryotube kusungitsa |
Madzi | Kwa zitsanzo zokhala ndi tizilombo tating'onoting'ono, monga madzi apampopi, madzi apachitsime, ndi zina zotero, Sonkhanitsani madzi osachepera 1 L ndikudutsa 0.22 μm fyuluta kuti mulemeretse tizilombo toyambitsa matenda pa nembanemba.Sungani nembanemba mu chubu chosabala. |
Khungu | Pala pamwamba pa khungu ndi thonje wosabala kapena tsamba la opaleshoni ndikuyika mu chubu wosabala. |
Ikani zitsanzo mu nayitrogeni wamadzimadzi kwa maola 3-4 ndikusunga mu nayitrogeni wamadzimadzi kapena -80 digiri kuti musungidwe nthawi yayitali.Kutumiza kwachitsanzo kokhala ndi ayezi wowuma ndikofunikira.
1.Histogram: Kugawa kwamitundu
2.Majini ogwira ntchito ofotokozera njira za metabolic za KEGG
Mapu a 3.Kutentha: Ntchito zosiyana zotengera kuchuluka kwa majini4.Circos of CARD antibiotic resistance genenes
BMK Case
Kuchuluka kwa majini olimbana ndi maantibayotiki ndi tizilombo toyambitsa matenda m'mphepete mwa mizu ya mangrove
Lofalitsidwa:Journal of Hazardous Equipment, 2021
Njira yotsatirira:
Zipangizo: Zotulutsa za DNA za zidutswa zinayi za mizu ya mangrove yokhudzana ndi zitsanzo: nthaka yosabzalidwa, rhizosphere, episphere ndi endosphere compartments
Pulatifomu: Illumina HiSeq 2500
Zolinga: Metagenome
16S rRNA jini V3-V4 dera
Zotsatira zazikulu
Tsatanetsatane wa metagenomic ndi mbiri ya metabarcoding pamizu yopitilira muzu wa mitengo ya mangrove adakonzedwa kuti aphunzire kufalitsa ma antibiotic resistance genes (ARGs) kuchokera kunthaka kupita ku zomera.Deta ya Metagenomic inavumbulutsa kuti 91.4% ya majini olimbana ndi maantibayotiki amadziwika bwino m'zigawo zonse zinayi za dothi zomwe zatchulidwa pamwambapa, zomwe zimasonyeza kachitidwe kosalekeza.16S rRNA amplicon sequencing adapanga 29,285 motsatira, kuyimira mitundu 346.Kuphatikizana ndi mbiri ya zamoyo pogwiritsa ntchito ma amplicon sequencing, kufalitsa kumeneku kunapezeka kuti sikunachokere ku microbiota yokhudzana ndi mizu, komabe, ikhoza kuthandizidwa ndi mafoni amtundu.Kafukufukuyu adazindikira kutuluka kwa ma ARGs ndi tizilombo toyambitsa matenda kuchokera m'nthaka kulowa muzomera kudzera munjira yolumikizana ndi mizu ya nthaka.
Buku
Wang, C. , Hu, R. , Strong, PJ , Zhuang, W. , & Shu, L. .(2020).Kuchuluka kwa majini olimbana ndi maantibayotiki ndi tizilombo toyambitsa matenda m'nthaka-mizu ya mangrove mosalekeza.Journal of Hazardous Materials, 408, 124985.