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Nā huahana

Ke Kaʻina Hana Metagenomic (NGS)

ʻO ka Metagenome e pili ana i ka hōʻiliʻili ʻana o nā ʻano genetic a pau o kahi kaiāulu hui o nā mea ola, e like me ka metagenome kaiapuni, metagenome kanaka, a pēlā aku.ʻO ka metagenomic sequencing he mea hana molecular i hoʻohana ʻia e kālailai i nā mea genomic hui ʻia i unuhi ʻia mai nā laʻana kaiapuni, e hāʻawi ana i ka ʻike kikoʻī i ka ʻano like ʻole a me ka nui o nā ʻano, ke ʻano o ka heluna kanaka, ka pilina phylogenetic, nā genes hana a me ka pilina pili me nā mea kaiapuni.

Papahana:Illumina NovaSeq6000


Nā kikoʻī lawelawe

Nā hopena hōʻike

Nānā Hiʻohiʻona

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

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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

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Hāʻawi laʻana

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Kūkulu hale waihona puke

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Ke kaʻina ʻana

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ʻIkepili ʻikepili

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Nā lawelawe ma hope o ke kūʻai aku


  • Mua:
  • Aʻe:

  • 1.Histogram: Hoʻolaha ʻano ʻano

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    2.Functional genes annotated to KEGG metabolic pathways

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    3. Heat map: Nā hana like ʻole e pili ana i ka nui o nā gene pili54.Circos o nā genes kū'ē antibiotic CARD

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    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.

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