Le fa'asinomaga fa'avae mRNA Sequencing-NGS
Vaega
● Pu'e le poly mRNA a'o le'i saunia le faletusi
● Tuto'atasi mai so'o se genome fa'asinomaga: fa'avae i luga o fa'apotopotoga de novo o tusitusiga, fa'atupuina se lisi o unigenes o lo'o fa'amatalaina i le tele o fa'amaumauga (NR, Swiss-Prot, COG, KOG, eggNOG, Pfam, GO, KEGG)
● Au'ili'iliga fa'aolaola o le fa'amatalaga o kenera ma le fa'asologa o tusitusiga
Tulaga Auaunaga
●Tomai Tele: fa'atasi ai ma se fa'amaumauga o le fa'agaioiina i luga ole 600,000 fa'ata'ita'iga ile BMKGENE, fa'ata'amilo i ituaiga fa'ata'ita'iga eseese e pei ole tu'ufa'atasiga o le cell, tissues, ma le sua o le tino, e aumaia e la tatou 'au le tele o le poto masani i galuega uma. Ua matou tapunia ma le manuia le silia ma le 100,000 mRNA-Seq poloketi i vaega eseese o suʻesuʻega.
●Pulea lelei: matou te fa'atinoina le fa'atonuga autu i la'asaga uma, mai le fa'ata'ita'iga ma tapenaga faletusi i le fa'asologa ma bioinformatics. O lenei mata'ituina lelei e fa'amautinoa ai le tu'uina atu o fa'ai'uga maualuluga.
● Faʻamatalaga Atoa: matou te fa'aogaina le tele o fa'amaumauga e fa'amatala ai le Differentially Expressed Genes (DEGs) ma fa'atino le su'esu'ega fa'atamaoaigaina talafeagai, tu'uina atu fa'amatalaga i luga o le telefoni feavea'i ma le molekalale o lo'o i lalo o le tali transcriptome.
●Lagolago Post-Fa'atau: o la matou tautinoga e faʻalautele atu i tua atu o le faʻamaeʻaina o galuega faʻatasi ma le 3-masina pe a maeʻa le faʻatau atu taimi auaunaga. I le taimi lea, matou te ofoina atu le tulitatao o poloketi, fesoasoani faʻafitauli, ma Q&A sauniga e faʻafeiloaʻi ai soʻo se fesili e fesoʻotaʻi ma iʻuga.
Fa'ata'ita'iga Mana'oga ma le Tu'uina atu
| Faletusi | Fuafuaga fa'asologa | Fa'amatalaga fautuaina | Puleaina lelei |
| Poly A fa'atamaoaigaina | Illumina PE150 DNSSEQ-T7 | 6-10 Gb | Q30≥85% |
Fa'ata'ita'iga Mana'oga:
Nucleotides:
| Conc.(ng/μl) | Aofa'i (μg) | Mama | Fa'amaoni |
| ≥ 10 | ≥ 0.2 | OD260/280=1.7-2.5 OD260/230=0.5-2.5 Fa'atapula'a pe leai se polotini po'o le fa'aleagaina o DNA o lo'o fa'aalia ile gel. | Mo laau toto: RIN≥4.0; Mo manu: RIN≥4.5; 5.0≥28S/18S≥1.0; fa'atapula'a pe leai se maualuga maualuga |
● La'au:
A'a, Po'o Po'o Petal: 450 mg
Laulaau po'o Fatu: 300 mg
Fualaau: 1.2 g
● Manu:
FOTO po'o le Tonu: 300 mg
Viscera po'o Fai'ai: 240 mg
Muso: 450 mg
Ivi, Laulu po'o Pa'u: 1g
● Arthropods:
Iniseti: 6g
Crustacea: 300 mg
● toto atoa: 1 paipa
● Selula: 106 sela
Fautuaina Fa'ata'ita'iga Tu'uina atu
Containe: 2 ml centrifuge tube (E le fautuaina le pepa apa)
Fa'ailoga fa'ata'ita'iga: Vaega+fa'atusa eg A1, A2, A3; B1, B2, B3.
utaina:
1. Fa'amamago-aisa: Fa'ata'ita'iga e mana'omia ona fa'apipi'i i taga ma tanu i le aisa-mago.
2. RNAstable tubes: RNA samples e mafai ona fa'amago ile RNA stabilization tube(eg RNAstable®) ma lafo ile potu vevela.
Auala o Galuega Au'aunaga
Fa'ata'ita'iga mamanu
Fa'ata'ita'iga tu'uina atu
RNA su'ega
Fausia faletusi
Fa'asologa
Iloiloga fa'amaumauga
Au'aunaga pe a uma ona fa'atau
Bioinformatics
Transcriptome fono ma unigene filifiliga
Unigene fa'amatalaga
Fa'ata'ita'iga feso'ota'iga ma su'esu'ega o fa'asologa o mea ola
Genes ua Fa'aalia Eseese (DEGs)
Fa'amatalaga Fa'atino o DEGs
Fa'atamaoaigaina Galuega ole DEGs
Su'esu'e le aga'iga'i i luma na fa'afaigofieina e le BMKGene's eukaryotic NGS mRNA sequencing services e ala i le aoina o fa'asalalauga.
Shen, F. et al. (2020) 'De novo transcriptome assembly and sex-biased gene expression in the gonads of Amur catfish (Silurus asotus)', Genomics, 112(3), itulau 2603–2614. doi: 10.1016/J.YGENO.2020.01.026.
Zhang, C. et al. (2016) 'Transcriptome analysis of sucrose metabolism i le taimi o le fulafula o le pulu ma le atinaʻeina o aniani (Allium cepa L.)', Frontiers in Plant Science, 7 (Setema), p. 212763. doi: 10.3389/FPLS.2016.01425/BIBTEX.
Zhu, C. et al. (2017) 'De novo faʻapotopotoga, faʻamatalaga ma faʻamatalaga mo le transcriptome o Sarcocheilichthys sinensis', PLoS ONE, 12(2). doi: 10.1371/JOURNAL.PONE.0171966.
Zou, L. et al. (2021) 'De novo transcriptome analysis e maua ai fa'amatalaga i le fa'apalepale o le masima o le Podocarpus macrophyllus i lalo o le mamafa o le salinity', BMC Plant Biology, 21(1), itulau 1–17. doi: 10.1186/S12870-021-03274-1/ATA/9.
