16S/18S/ITS Amplicon Sequencing-PacBio
Pono lawelawe
● Heluhelu lōʻihi e hōʻike ana i ke kaʻina holoʻokoʻa o 16S/18S/ITS
● Kāhea kumu pololei loa me ke kaʻina o ke ʻano CCS PacBio
● ʻO ka hoʻonā ʻano ʻano-level ma ka hōʻike OTU/ASV
● ʻO ke kahe anana QIIME2 hou loa me nā ʻano kālailai like ʻole e pili ana i ka waihona waihona, hōʻike, OTU/ASV.
● Pili i nā haʻawina kaiaulu microbial like ʻole
● Loaʻa iā BMK ka ʻike nui me ka ʻoi aku o 100,000 mau laʻana / makahiki， uhi ʻia ka lepo, ka wai, ke kinoea, ka ʻōpala, nā feces, nā ʻōpū, ka ʻili, ka hupa fermentation, nā inike, nā mea kanu, etc.
● Ua maʻalahi ʻo BMKCloud i ka wehewehe ʻikepili i loaʻa iā 45 mau mea hana loiloi pilikino
Nā kikoʻī lawelawe
| Ke kaʻina ʻanaPapahana | Hale Waihona Puke | Manaʻo ʻikepili | Ka manawa huli |
| PacBio Sequel II | SMRT-bele | 5K/10K/20K mau huaʻōlelo | 44 Nā lā hana |
Nā loiloi bioinformatics
● Ka hoʻomalu maikaʻi ʻikepili maka
● OTU clustering/De-noise(ASV)
● palapala hōʻike OTU
● ʻokoʻa Alpha
● ʻokoʻa Beta
● Ka nānā ʻana ma waena o ka hui
● Kānāwai hui kūʻē i nā kumu hoʻokolohua
● wānana gene hana
Nā Koina Laʻana a me ka Hoʻouna ʻana
Nā Koina Laʻana:
No ka meaNā unuhi DNA:
| ʻAno Laʻana | Ka nui | Hoʻopaʻa ʻana | Maemae |
| Nā unuhi DNA | ＞ 1 μg | ＞ 20 ng/μl | OD260/280= 1.6-2.5 |
No nā laʻana kaiapuni:
| ʻAno laʻana | Manaʻo ʻia ke kaʻina hana laʻana |
| lepo | Ka nui o ka laʻana: approx.5 g;Pono e wehe ʻia nā mea maloʻo i koe mai ka ʻili;E wili i nā ʻāpana nui a hele i ka kānana 2 mm;ʻO nā laʻana Aliquot i loko o ka sterile EP-tube a i ʻole cyrotube no ka mālama ʻana. |
| Feces | Ka nui o ka laʻana: approx.5 g;E hōʻiliʻili a hāʻawi i nā laʻana i loko o ka sterile EP-tube a i ʻole cryotube no ka mālama ʻana. |
| ʻO loko o ka ʻōpū | Pono e hana ʻia nā laʻana ma lalo o ke kūlana aseptic.Holoi i ka ʻiʻo i hōʻiliʻili ʻia me ka PBS;Centrifuge i ka PBS a ohi i ka precipitant ma EP-tubes. |
| Sludge | Ka nui o ka laʻana: approx.5 g;E hōʻiliʻili a hāʻawi i kahi laʻana o ka lepo i loko o ka EP-tube sterile a i ʻole cryotube no ka mālama ʻana |
| Ke kino wai | No ka laʻana me ka liʻiliʻi o ka microbial, e like me ka wai paipu, wai punawai, a me nā mea ʻē aʻe.E mālama i ka membrane i loko o ka paipu sterile. |
| ʻili | E ʻoki pono i ka ʻili o ka ʻili me ka pulupulu maʻemaʻe a i ʻole ka lāʻau lapaʻau a waiho i loko o ka paipu sterile. |
Manaʻo ʻia ka hāʻawi laʻana
E hoʻokuʻu i nā laʻana i loko o ka hau hau no 3-4 mau hola a e mālama i loko o ka wai nitrogen a i ʻole -80 degere a hiki i ka mālama lōʻihi.Pono ka hoʻouna laʻana me ka hau maloʻo.
Holo Hana Hana
Hāʻawi laʻana
Kūkulu hale waihona puke
Ke kaʻina ʻana
ʻIkepili ʻikepili
Nā lawelawe ma hope o ke kūʻai aku
1. Ka helu hoʻolaha o V3+V4(Illumina)-based microbial community profiling vs. Full-longth (PacBio)-based profiling.
(Ua noi ʻia nā ʻikepili o 30 mau papahana i koho ʻole ʻia no ka ʻikepili)
2.Annotation rate of full-length amplicon sequencing at species-level ma nā ʻano hāpana like ʻole
3. Hoʻolaha ʻano ʻano
4.Phylogenetic lāʻau
Hihia BMK
Hoʻopuka ka Arsenic i ka pōʻino o ka ʻōpū a me ka hoʻōla ʻana o ka axis gut-liver e alakaʻi ai i ka mumū a me ka pyroptosis o ke ake i loko o nā pā.
Paʻi ʻia:ʻepekema o ke kaiapuni holoʻokoʻa,2021
Hoʻolālā kaʻina hana:
Nā Laʻana: Mana vs 8 mg/kg ATO pūʻulu hōʻike
Ka helu ʻana i ka ʻikepili: 102,583 mau kaʻina CCS maka i ka huina
Mana: 54,518 ± 747 CCS pono
Hōʻike ʻia ʻo ATO : 45,050 ± 1675 CCS kūpono
Nā hopena koʻikoʻi
ʻokoʻa Alpha:Ua hoʻololi nui ka hōʻike ʻana iā ATO i ka waiwai microbial intestinal a me ka ʻokoʻa i loko o nā ʻeke.
ʻIkepili metastats:
I ka pae phylum: 2 phyla bacteria i ʻike wale ʻia ma nā pūʻulu mana
I ka pae genus: 6 mau genera i ʻike ʻia he ʻokoʻa loa i ka nui pili
I ka pae ʻano: 36 mau ʻano i ʻike ʻia i ka huina, me 6 o lākou he ʻokoʻa loa i ka nui o ka hoʻōla.
Kuhikuhi
Thingholm, LB , et al."ʻO ka poʻe obese me ka maʻi maʻi type 2 a me ka ʻole o ka maʻi diabetes e hōʻike i ka mana a me ka hoʻokumu ʻana o ka ʻāʻī microbial."Pūnaehana Pūnaewele & Microbe26.2(2019).
