16S/18S/ITS Amplicon Sequencing-PacBio
Amfanin Sabis
● Dogon karantawa yana bayyana cikakken jerin jerin 16S/18S/ITS
● Madaidaicin kiran tushe tare da tsarin yanayin PacBio CCS
● Ƙirar-matakin nau'i a cikin bayanin OTU/ASV
● Sabbin QIIME2 na nazarin kwarara tare da nazari iri-iri dangane da bayanan bayanai, annotation, OTU/ASV.
● 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 | Bayanan da aka ba da shawarar | Lokacin Juya |
| PacBio Sequel II | SMRT - kararrawa | 5K/10K/20K Tags | 44 Kwanakin aiki |
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 | 1 μg | 20 ng/ml | 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. Ƙididdigar bayanin V3+V4 (Illumina) - tushen ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun bayanai (PacBio) - tushen bayanin martaba.
(An yi amfani da bayanan ayyukan 30 da aka zaɓa don ƙididdiga)
2. Annotation rate na cikakken tsawon amplicon sequencing a jinsi-matakin a daban-daban samfurin iri
3.Species rarraba
4.Bishiyar phylogenetic
BMK Case
Bayyanar cututtuka na arsenic yana haifar da lalacewar shinge na hanji da sakamakon kunnawa na gut-hanta axis wanda ke haifar da kumburi da pyroptosis na hanta a cikin ducks.
Buga:Kimiyya na Jumlar Muhalli,2021
Dabarun tsarawa:
Samfurori: Sarrafa vs 8 mg/kg ATO fallasa kungiyar
Adadin bayanan jeri: 102,583 danyen jerin CCS gabaɗaya
Sarrafa: 54,518 ± 747 tasiri CCS
An fallasa ATO: 45,050 ± 1675 ingantaccen CCS
Sakamako mai mahimmanci
Bambancin Alpha:Bayyanar ATO sosai ya canza wadatar ƙananan ƙwayoyin cuta na hanji da bambancin a cikin agwagwa.
Binciken Metastats:
A matakin phylum: 2 kwayoyin phyla kawai an gano su a cikin ƙungiyoyi masu sarrafawa
A cikin Heart Level: 6 Genera aka samo mahimmanci a cikin dangi da yawa
A cikin nau'in halitta: An gano nau'ikan 36 a cikin duka, tare da 6 daga gare su daban-daban cikin yawa
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).
