ʻO ke Kaʻina Metagenomic-Nanopore
Pono lawelawe
● Kiʻekiʻe kiʻekiʻe-e hoʻonui i ka pololei o ka ʻike ʻana i nā ʻano a me ka wānana gene funcational
● Paʻa ka hoʻokaʻawale genome bacteria
● Hoʻohana ikaika a hilinaʻi hoʻi ma nā wahi like ʻole, e laʻa me ka ʻike ʻana i nā microorganism pathogenic a i ʻole nā genes pili i ka pale antibiotic.
● Ke kālailai metagenome hoʻohālikelike
Nā kikoʻī lawelawe
| Papahana | Ke kaʻina ʻana | Manaʻo ʻikepili | Ka manawa huli |
| Nanopore | ONT | 6 G/10 G | 65 mau lā hana |
Nā loiloi bioinformatics
● Ka mana o ka maikaʻi o ka ʻikepili maka
● Hui Metagenome
● ʻAʻole-redundant gene set and annotation
● Ka nānā 'ana i nā 'ano like 'ole
● Ke kālailai ʻokoʻa hana genetic
● Ka nānā ʻana ma waena o ka hui
● Kānāwai hui kūʻē i nā kumu hoʻokolohua
Nā Koina Laʻana a me ka Hoʻouna ʻana
Nā koi laʻana a me ka lawe ʻana
Nā Koina Laʻana:
No ka meaNā unuhi DNA:
| ʻAno Laʻana | Ka nui | Hoʻopaʻa | Maemae |
| Nā unuhi DNA | 1-1.5 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, ka wai punawai, a me nā mea ʻē aʻe, E hōʻiliʻili i ka liʻiliʻi o 1 L wai a hele i ka kānana 0.22 μm e hoʻonui i ka microbial ma ka membrane.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 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. Heatmap: Hui waiwai waiwai
2.Functional genes annotated to KEGG metabolic pathways
3.Species correlation network
4.Circos o CARD antibiotic kū'ē genes
Hihia BMK
Hiki i ka Nanopore metagenomics ke hōʻoia wikiwiki i ka maʻi bacterial haʻahaʻa hanu
Paʻi ʻia:Nature Biotechnology, 2019
Nā Manaʻo Kūikawā
Ka hoʻonohonoho ʻana: Nanopore Minion
ʻO nā bioinformatics metagenomics maʻi: Hoʻopau i ka DNA Host, WIMP a me ka loiloi ARMA
ʻIke wikiwiki: 6 hola
ʻIke kiʻekiʻe: 96.6%
Nā hopena koʻikoʻi
I ka makahiki 2006, ʻo ka maʻi hanu haʻahaʻa (LR) ka mea i make i 3 miliona mau kānaka ma ka honua holoʻokoʻa.ʻO ke ʻano maʻamau no ka ʻike ʻana i ka pathogen LR1, ʻo ia ka mahiʻai, ʻaʻohe naʻau, lōʻihi ka huli ʻana a ʻaʻohe alakaʻi i ka lāʻau antibiotic mua.ʻO ka maʻi microbial wikiwiki a pololei i ka manawa lōʻihi he pono koʻikoʻi.Ua hoʻomohala maikaʻi ʻo Kauka Justin mai ke Kulanui o East Anglia a me kāna mau hoa hana i kahi ʻano metagenomic e pili ana i Nanopore no ka ʻike pathogen.Wahi a kā lākou kaʻina hana, hiki ke hoʻopau ʻia ka 99.99% o ka DNA host.Hiki ke hoʻopau ʻia ka ʻike ʻana i nā pathogens a me nā genes kūpaʻa antibiotic i 6 mau hola.
Kuhikuhi
Charalampous, T. , Kay, GL , Richardson, H. , Aydin, A. , & O'Grady, J. .(2019).Hiki i ka Nanopore metagenomics ke hōʻoia wikiwiki i ka maʻi bacterial haʻahaʻa hanu.Nature Biotechnology, 37(7), 1.
