Metagenomic Sequencing-Nanopore
Tulaga Auaunaga
● Fa'apotopotoga maualuga-Fa'aleleia le sa'o o le fa'ailoaina o ituaiga ma va'aiga gene fa'aoga
● Tapuni le fa'aesea o le genome siama
● E sili atu le malosi ma le fa'atuatuaina o le fa'aogaina i vaega eseese, fa'ata'ita'iga, su'esu'eina o meaola ninii fa'ama'i po'o kenera e tete'e i antibiotic.
● Su'esu'ega metagenome fa'atusatusa
Fa'amatalaga Auaunaga
| Fa'avae | Fa'asologa | Fa'amatalaga fautuaina | Taimi Fa'aliliu |
| Nanopore | ONT | 6 G/10 G | 65 Aso faigaluega |
Bioinformatics su'esu'ega
● Pulea lelei o fa'amaumauga mata'utia
● Fonotaga Metagenome
● Seti o kenera e le toe faia ma fa'amatalaga
● Su'esu'ega o ituaiga eseese
● Su'esu'ega o galuega fa'atupu kenera
● Su'esu'ega va'aiga
● Su'esu'ega fa'atasi e fa'atatau i mea fa'ata'ita'i
Fa'ata'ita'iga Mana'oga ma le Tu'uina atu
Faʻataʻitaʻiga manaʻoga ma tuʻuina atu
Fa'ata'ita'iga Mana'oga:
MoDNA extracts:
| Ituaiga Fa'ata'ita'i | Aofaiga | Fa'atotonuga | mama |
| DNA extracts | 1-1.5 μg | + 20 ng/μl | OD260/280= 1.6-2.5 |
Mo fa'ata'ita'iga tau si'osi'omaga:
| Fa'ata'ita'iga ituaiga | Fautuaina faiga faataitai |
| Eleele | Aofa'i fa'ata'ita'iga: tusa.5 g;O mea maguma totoe e tatau ona aveese mai luga;Oli fasipepa lapopoa ma pasi i totonu ole faamama 2 mm;Aliquot faʻataʻitaʻiga i le sterile EP-tube poʻo le cyrotube mo le faʻaagaga. |
| Feces | Aofa'i fa'ata'ita'iga: tusa.5 g;Aoina ma fa'ata'ita'i fa'ata'ita'iga ile EP-tube po'o le cryotube mo le fa'aagaga. |
| I totonu o le manava | E mana'omia le fa'agaioiina o fa'ata'ita'iga i lalo ole tulaga aseptic.Fufulu aano o lo'o aoina ile PBS;Centrifuge le PBS ma fa'aputu le mea fa'afefe ile EP-tubes. |
| Sludge | Aofa'i fa'ata'ita'iga: tusa.5 g;Aoina ma fa'ata'ita'i fa'ata'ita'i palapala ile EP-tube po'o le cryotube mo le fa'aagaga |
| Vaivai | Mo faʻataʻitaʻiga e faʻatapulaʻa le aofaʻi o microbial, e pei o le paipa vai, vai vai, ma isi, Aʻoaʻo mai le itiiti ifo i le 1 L vai ma pasi ile 0.22 μm faamama e faʻatamaoaigaina ai microbial i luga ole membrane.Teu le pa'u i totonu o le faagaau mama. |
| Pa'u | 'A'e ma le fa'aeteete le pa'u i le su'e fugala'au po'o le lau tipitipi ma tu'u i totonu ole faagaau mama. |
Fautuaina Fa'ata'ita'iga Tu'uina atu
Fa'a'aisa fa'ata'ita'iga ile sua vai mo le 3-4 itula ma teu ile fa'asusu vai po'o le -80 tikeri ile fa'aagaga umi.Fa'ata'ita'iga felauaiga fa'atasi ai ma le aisa mago e mana'omia.
Auala o Galuega Au'aunaga
Fa'ata'ita'iga tu'uina atu
Fausia faletusi
Fa'asologa
Iloiloga fa'amaumauga
Au'aunaga pe a uma ona fa'atau
1. Heatmap: Fa'aputuina o'oa o ituaiga
2. Functional genes annotated to KEGG metabolic pathways
3.Species feso'ota'iga feso'ota'iga
4.Circos of CARD antibiotic resistance genes
BMK mataupu
Nanopore metagenomics e mafai ai ona vave suʻesuʻeina faʻamaʻi o siama faʻamaʻi maualalo manava
Lolomiina:Natura Biotechnology, 2019
Fa'amatalaga Fa'apitoa
Fa'asologa: Nanopore Minion
Bioinformatics metagenomics ile falemaʻi: Faʻaleagaina DNA faʻapitoa, suʻesuʻega WIMP ma ARMA
Saili vave: 6 itula
Ma'ale'ale maualuga: 96.6%
I'uga autu
I le 2006, na mafua ai le 3 miliona tagata maliliu i le lalolagi atoa.O le auala masani mo le su'esu'eina o le pathogen LR1 o le fa'ato'aga, lea e leaga le fa'alogona, umi le taimi e liliu ai ma leai se ta'iala i togafitiga fa'ama'i vave.Ole su'esu'ega vave ma le sa'o ole microbial ua leva ona avea ma mana'oga fa'anatinati.Dr. Justin mai le Iunivesite o East Anglia ma ana pa'aga na fa'atupuina ma le manuia se metotia metagenomic fa'avae Nanopore mo le su'esu'eina o pathogen.E tusa ai ma la latou galuega, 99.99% o le DNA talimalo e mafai ona faʻaumatia.E mafai ona fa'amae'a ile 6 itula ole su'esu'ega ile siama ole ma'i fa'ama'i.
Fa'asinomaga
Charalampous, T. , Kay, GL , Richardson, H. , Aydin, A. , & O'Grady, J. .(2019).Nanopore metagenomics e mafai ai ona vave suʻesuʻeina faʻamaʻi o siama faʻamaʻi maualalo manava.Natura Biotechnology, 37(7), 1.
