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16S/18S/ITS Amplicon Sequencing-PacBio

Kwayoyin halittar 16S da 18S rRNA, tare da yankin Internal Transcribed Spacer (ITS), suna aiki a matsayin alamomin zanen yatsa na kwayoyin halitta saboda hadewarsu na yankuna da aka kiyaye sosai da masu saurin canzawa, suna sanya su kayan aiki masu mahimmanci don siffanta kwayoyin prokaryotic da eukaryotic. Ƙwaƙwalwa da jeri na waɗannan yankuna suna ba da hanyar da ba ta da keɓancewa don bincika ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun yanayin halittu daban-daban. Yayin da jerin abubuwan Illumina yawanci ke kaiwa ga gajerun yankuna masu canzawa kamar V3-V4 na 16S da ITS1, an nuna cewa ana iya samun ingantaccen bayanin haraji ta hanyar yin cikakken tsayin 16S, 18S, da ITS. Wannan cikakkiyar dabarar tana haifar da mafi girman kashi na daidaitattun jeri-jeri, cimma matakin ƙuduri wanda ya kai ga tantance nau'in. PacBio's Single-Molecule Real-Time Real-Time (SMRT) dandali na jeri ya fice ta hanyar samar da ingantaccen dogon karantawa (HiFi) wanda ke rufe cikakken amplicons, yana fafatawa da madaidaicin jerin Illumina. Wannan damar tana ba masu bincike damar samun fa'ida mara misaltuwa - ra'ayi mai ban mamaki game da yanayin yanayin halitta. Tsawaita ɗaukar hoto yana haɓaka ƙuduri sosai a cikin bayanin nau'ikan, musamman a cikin al'ummomin ƙwayoyin cuta ko na fungal, yana ba da damar fahimtar zurfafa fahimtar rikice-rikice na yawan ƙananan ƙwayoyin cuta.

Tuntube Mu

Cikakkun Sabis

Bioinformatics

Sakamakon Demo

Fitattun wallafe-wallafe

Siffofin Sabis

● Tsarin dandali: PacBio Revio

● Yanayin jeri: CCS (Karanta HiFi)

● Ƙaddamar da yankin da aka yi niyya tare da haɗin kai na amplicons kafin shirye-shiryen ɗakin karatu na HiFi SMRT

Amfanin Sabis

●Ƙididdigar haraji mafi girma: Than short-amplicon jerin,ba da damar mafi girman ƙimar rarrabuwar OTU a matakin nau'in.

●Ingantacciyar kiran tushe: Tsarin yanayin PacBio CCS (Karanta HiFi).

●Babu warewa: Saurin gano ƙwayoyin ƙwayoyin cuta a cikin samfuran muhalli.

●Yadu aiki: Nazarin al'umma daban-daban na ƙananan ƙwayoyin cuta.

●Cikakken bincike na bioinformatic: Sabbin fakitin QIIME2 (ƙididdigar ƙididdigewa game da ilimin halittu) tare da nazari iri-iri dangane da bayanan bayanai, annotation, OTU/ASV.

●Ƙwararren Ƙwararru: Tare da dubban ayyukan jerin amplicon da aka gudanar a kowace shekara, BMKGENE yana kawo shekaru goma na kwarewa, ƙungiyar bincike mai ƙwarewa, cikakkun abun ciki, da kuma kyakkyawan goyon bayan tallace-tallace.

Ƙayyadaddun Sabis

Laburare	Dabarun Jeri	An ba da shawarar bayanai
Amplicon	PacBio Revio	10/30/50 K tags (CCS)

Samfuran Bukatun

Hankali (ng/µL)	Jimlar adadin (µg)	girma (µL)
≥5	≥0.3	≥20

Isar da Samfurin Nasiha

Daskare samfuran a cikin ruwa nitrogen na sa'o'i 3-4 kuma adana a cikin ruwa nitrogen ko -80 digiri zuwa ajiyar dogon lokaci. Ana buƙatar jigilar samfuri tare da bushe-kankara.

Gudun Aikin Sabis

Samfurin bayarwa

Gina ɗakin karatu

Jeri

Binciken bayanai

Bayan-sayar da sabis

Na baya: 16S/18S/ITS Amplicon Sequencing-NGS

Na gaba: Ƙididdiga don Canjawa-Sarfafa Chromatin tare da Babban Sabis na Sabis (ATAC-seq)

Ya haɗa da bincike mai zuwa:

●Raw data ingancin kula

●OTU clustering/De-noise(ASV)

● Bayanin OTU

● Binciken bambancin Alpha: ma'auni masu yawa, ciki har da Shannon, Simpson da ACE.

●Binciken bambancin beta

●Bincike tsakanin ƙungiyoyi

●Binciken daidaitawa: tsakanin abubuwan muhalli da abubuwan OUT da bambancin

● 16S aikin tsinkayar kwayoyin halitta

Histogram na rarraba taxonomic

Al'umma rarraba phylogenetic itace

Binciken bambancin Alpha: ACE

index

Binciken bambancin Beta: PCoA

Binciken Intergroup: ANOVA

Bincika ci gaban da sabis na amplicon na BMKGene ya sauƙaƙe tare da PacBio ta hanyar tarin wallafe-wallafe.

Gao, X. da Wang, H. (2023) 'Comparative Analysis of Rumen Bacterial Profiles and Services while Adaption to Daban-daban Phenology (Regreen vs. Grassy) a cikin Alpine Merino Sheep tare da Matakan Girma Biyu a kan Dutsen Mai tsayi', Fermentation, 9 ( 1), p. 16. doi: 10.3390/FERMENTATION9010016/S1.

Li, S. et al. (2023) 'Kwaƙwalwar ƙwayoyin cuta masu duhu a cikin ƙasa hamada ta amfani da culturomics-tushen metagenomics da high-ƙuduri bincike', npj Biofilms da Microbiomes 2023 9: 1, 9 (1), shafi 1-14. doi: 10.1038/s41522-023-00439-8.

Mu, L. et al. (2022) 'Sakamakon m acid salts a kan fermentation halaye, na kwayan cuta bambancin da aerobic kwanciyar hankali na gauraye silage shirya tare da alfalfa, shinkafa bambaro da alkama bran', Journal of the Science of Food and Agriculture, 102(4), shafi na 1475- 1487. doi: 10.1002/JSFA.11482.

Yang, J. et al. (2023) 'Haɗin kai tsakanin Oxidative Stress Biomarkers da Gut Microbiota a cikin Tasirin Antioxidant na Cire daga Sonchus brachyotus DC. a cikin Oxazolone-Induced Intestinal Oxidative stress in Adult Zebrafish', Antioxidants 2023, Vol. 12, Shafi na 192, 12 (1), shafi. 192. doi: 10.3390/ANTIOX12010192.