Kutsata kwa Metagenomic-TGS
Metagenome ndi gulu la majini a gulu losakanikirana la zamoyo, monga ma metagenome achilengedwe ndi anthu. Lili ndi majini a tizilombo tomwe tingalimidwe komanso tosalimidwa. Kutsata kwa metagenomic kumathandiza kuphunzira malo ovuta awa a genomic omwe ali mu zitsanzo za chilengedwe popereka zambiri kuposa kutsata kwa taxonomic. Kumaperekanso lingaliro logwira ntchito la genomics pofufuza majini olembedwa ndi ntchito zawo zoganizira muzochitika zachilengedwe. Ngakhale njira zachikhalidwe zogwiritsira ntchito Illumina sequencing zakhala zikugwiritsidwa ntchito kwambiri mu maphunziro a metagenomic, kubwera kwa Nanopore ndi PacBio sequencing kwasintha munda. Ukadaulo wa Nanopore ndi PacBio umathandizira kusanthula kwa bioinformatic pansi, makamaka kusonkhana kwa metagenome, kuonetsetsa kuti makonzedwe opitilira. Malipoti akusonyeza kuti ma metagenomics ochokera ku Nanopore ndi PacBio apanga bwino ma genome athunthu komanso otsekedwa a bakiteriya kuchokera ku ma microbiomes ovuta (Moss, EL, et al., Nature Biotech, 2020). Kuphatikiza ma Nanopore reads ndi ma Illumina reads kumapereka njira yabwino yokonzera zolakwika, kuchepetsa kulondola kochepa kwa Nanopore. Kuphatikizana kumeneku kumagwiritsa ntchito mphamvu za nsanja iliyonse yotsatizana, kupereka yankho lolimba kuti ligonjetse zofooka zomwe zingatheke ndikupititsa patsogolo kulondola ndi kudalirika kwa kusanthula kwa metagenomic.
Nsanja: Nanopore PromethION 48, Illumia ndi PacBio Revio
Zinthu Zofunika pa Utumiki
● Kuwerenga kwa nthawi yayitali pa Nanopore P48 (laibulale ya 10Kb); PacBio Revio (laibulale ya 8Kb)
● Kukonza zolakwika: kutsatana kwa Illumina NovaSeq (laibulale ya PE150)
Ubwino wa Utumiki
●Msonkhano Wapamwamba wa Metagenome:Ndi ma contig ochepa omwe ali ndi continuity yapamwamba.
●Kulondola Kwambiri:Kuzindikira mitundu ndi kulosera za majini ogwira ntchito.
●Kukonza Koyenera: Kuthandizira kusiyanitsa majini a bakiteriya ndi kusanthula koyerekeza kwa metagenome.
●Kusanthula Kwathunthu kwa Bioinformatics:Kusanthula kwathu sikungoyang'ana pa kusiyanasiyana kwa taxonomic kokha komanso pa kusiyanasiyana kwa ntchito za anthu ammudzi.
●Chidziwitso ChambiriGulu lathu limabweretsa chidziwitso chochuluka ku projekiti iliyonse, ndi mbiri yabwino yomaliza bwino mapulojekiti angapo a metagenomics m'magawo osiyanasiyana ofufuza ndikugwiritsa ntchito zitsanzo zoposa 200,000, gulu lathu limabweretsa chidziwitso chochuluka ku projekiti iliyonse.
Mafotokozedwe a Utumiki
| Nsanja yotsatizana | Njira Yotsatizana | Deta ikulangizidwa |
| Illumina NovaSeq | PE150 | 6 Gb |
| Nanopore P48 | 10 kb | 6 / 10 Gb |
| PacBio Revio | 8 kb | 10 / 20 Gb |
Zofunikira pa Zitsanzo
| Kuchuluka kwa madzi (ng/µL) | Chiwerengero chonse (µg) | Voliyumu (µL) | |
| Laibulale ya Illumina PE150 | ≥1 | ≥0.03 | ≥20 |
| Laibulale ya Nanopore 10 kb | ≥40 | ≥2 | ≥20 |
| Laibulale ya PacBio 8 kb | ≥50 | 10 µg/selo | ≥20 |
Kayendedwe ka Ntchito Yotumikira
Kutumiza zitsanzo
Kumanga laibulale
Kutsatana
Kusanthula deta
Ntchito zogulitsa pambuyo pogulitsa
Kusanthula kotsatiraku kumaphatikizapo:
● Kuwongolera Ubwino wa Deta Motsatizana
● Kusonkhanitsa Metagenome ndi Kuneneratu za Majini
● Chidziwitso cha Majini
● Kusanthula kwa Mitundu ya Alpha ya Taxonomic
● Kusanthula Ntchito za Anthu: Ntchito ya Zamoyo, Kagayidwe kachakudya, Kukana Mankhwala Opha Ululu
● Kusanthula pa Kusiyanasiyana kwa Ntchito ndi Taxonomic:
Kusanthula kwa Mitundu Yosiyanasiyana ya Beta
Kusanthula kwa magulu osiyanasiyana
Kusanthula kwa Ubale: pakati pa zinthu zachilengedwe ndi kapangidwe ndi kusiyanasiyana kwa OUT
Kugawa kwa majini osagwiritsidwa ntchito mopitirira muyeso:
Chidziwitso cha majini: Njira ya KEGG
Chidziwitso cha jini: eggNOG
Chidziwitso cha majini: Ma carbohydrate osavuta
Ulalo wa mitundu:
Fufuzani kupita patsogolo komwe kwathandizidwa ndi ntchito ya BMKGene yofufuza metagenomics pogwiritsa ntchito Nanopore + Illumina ndi buku ili:
Jin, H. ndi ena. (2023) 'Kuphatikiza kwapamwamba kwa majini a m'mimba mwa munthu kwa anthu amkati mwa Mongolia', Nature Microbiology 2023 8:1, 8(1), pp. 150–161. doi: 10.1038/s41564-022-01270-1.
