Ke Kaʻina Hana Metagenomic (NGS)
Nā Pono lawelawe
ØʻO ka hoʻokaʻawale ʻana a me ka mahiʻai ʻole no ka profiling kaiāulu microbial
ØʻO ka hoʻonā kiʻekiʻe i ka ʻimi ʻana i nā ʻano haʻahaʻa haʻahaʻa i nā laʻana kaiapuni
ØHoʻohui ka manaʻo o "meta-" i nā hiʻohiʻona olaola āpau ma ka pae hana, ka pae ʻano a me ka pae gene, e hōʻike ana i kahi hiʻohiʻona ikaika e pili kokoke ana i ka ʻoiaʻiʻo.
ØHoʻonui ʻo BMK i ka ʻike nui ma nā ʻano laʻana like ʻole me ka ʻoi aku o 10,000 mau laʻana i hana ʻia.
Nā kikoʻī lawelawe
| Ke kaʻina ʻanaPapahana | Hale Waihona Puke | Hāʻawi ʻia ka ʻikepili i manaʻo ʻia | Kuhi ʻia ka manawa huli |
| ʻO Illumina NovaSeq 6000 | PE250 | 50K/100K/300K mau huaʻōlelo | 30 mau lā |
Nā loiloi bioinformatics
üKa mana maikaʻi ʻikepili maka
üHui metagenome
üʻO ka hoʻonohonoho gene a me ka wehewehe ʻole
üKa nānā 'ana i nā 'ano like 'ole
üKe kālailai ʻokoʻa hana genetic
üʻIkepili waena
üKa hoʻopili ʻana o ka hui kūʻē i nā mea hoʻokolohua
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 | ＞ 30 ng | ＞ 1 ng/μl | OD260/280= 1.6-2.5 |
No nā laʻana kaiapuni:
| ʻAno laʻana | Manaʻo ʻia ke kaʻina hana laʻana |
| Ka 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 hōʻiliʻili i ka precipitant ma EP-tubes. |
| Sludge | Ka nui o ka laʻana: approx.5 g;E hōʻiliʻili a hāʻawi i kahi laʻana sludge i loko o ka sterile EP-tube 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 akahele 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
Hoʻokuʻu i nā laʻana i loko o ka wai 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.Histogram: Hoʻolaha ʻano ʻano
2.Functional genes annotated to KEGG metabolic pathways
3. Heat map: Nā hana like ʻole e pili ana i ka nui o nā gene pili
4.Circos o nā genes kū'ē antibiotic CARD
Hihia BMK
ʻO ka laha ʻana o nā genes kūpaʻa antibiotic a me nā pathogens bacteria ma ka hoʻomau o ka lepo-mangrove root continuum
Paʻi ʻia:Nūpepa o nā Mea Pilikino, 2021
Hoʻolālā kaʻina hana:
Nā mea waiwai: Nā ʻāpana DNA o nā ʻāpana ʻehā o nā kumu lāʻau mangrove e pili ana: lepo kanu ʻole, rhizosphere, episphere a me nā keʻena endosphere.
Papahana: Illumina HiSeq 2500
Nā pahuhopu: Metagenome
16S rRNA gene V3-V4 māhele
Nā hopena koʻikoʻi
Ua hana ʻia ke kaʻina hana metagenomic a me ka metabarcoding profiling ma ka ʻaʻa lepo o nā kumu mangrove i mea e aʻo ai i ka hoʻolaha ʻana o nā genes resistance antibiotic (ARGs) mai ka lepo i nā mea kanu.Ua hōʻike ʻia ka ʻikepili Metagenomic he 91.4% o nā genes resistance antibiotic i ʻike maʻamau i loko o nā keʻena lepo ʻehā i ʻōlelo ʻia ma luna, i hōʻike ʻia i ke ʻano mau.16S rRNA amplicon sequencing i hana 29,285 sequencing, e hōʻike ana i 346 ʻano.ʻO ka hui pū ʻana me ka hoʻopili ʻana i nā ʻano mea e ka amplicon sequencing, ua ʻike ʻia kēia hoʻolaha ʻana he kūʻokoʻa i ka microbiota pili i ke aʻa, akā naʻe, hiki ke hoʻomaʻamaʻa ʻia e ka mobile o nā mea genetic.Ua ʻike kēia haʻawina i ka kahe ʻana o nā ARG a me nā pathogens mai ka lepo i loko o nā mea kanu ma o ka hoʻomau ʻana o ka lepo-aʻa.
Kuhikuhi
Wang, C. , Hu, R. , Strong, PJ , Zhuang, W. , & Shu, L. .(2020).ʻO ka laha ʻana o nā genes kūpaʻa antibiotic a me nā maʻi bacterial ma ka lepo-aʻa kumu mangrove.Nūpepa o nā mea pōʻino, 408, 124985.
