Fa'asologa o tusitusiga atoa - Illumina
Vaega
● Faletusi e lua e fa'asolo ai le fa'asologa atoa: fa'aitiitiga o le rRNA soso'o ma le tapenaga o le faletusi PE150 ma le lapopo'a fa'asolo ma le tapenaga o le potutusi SE50.
● Fa'auma su'esu'ega bioinformatics o le mRNA, lncRNA, circRNA ma le miRNA i lipoti eseese bioinformatics
● Su'esu'ega fa'atasi o fa'amatalaga RNA uma i lipoti tu'ufa'atasi, e aofia ai su'esu'ega o feso'ota'iga ceRNA.
Tulaga Auaunaga
●Iloiloga loloto o fesoʻotaʻiga faʻatonutonu: o le su'esu'ega o feso'ota'iga ceRNA e mafai ona fa'aogaina i le fa'asologa fa'atasi o le mRNA, lncRNA, circRNA ma le miRNA fa'atasi ai ma se fa'agasologa ole bioinformatic vaivai.
●Fa'amatalaga Atoa: matou te fa'aogaina le tele o fa'amaumauga e fa'ailoa ai fa'amatalaga o le Differentially Expressed Genes (DEGs) ma fa'atino le su'esu'ega fa'atamaoaigaina talafeagai, e maua ai fa'amatalaga i luga o le cellular ma molecular process o lo'o fa'avaeina le tali transcriptome.
●Tomai Tele: fa'atasi ai ma se fa'amaumauga o le tapunia manuia o le silia ma le 2000 galuega fa'akomepiuta atoa i vaega su'esu'e 'ese'ese, e aumaia e le matou 'au le tele o le poto masani i galuega uma.
●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'ailoa ai fa'amatalaga o le Differentially Expressed Genes (DEGs) ma fa'atino le su'esu'ega fa'atamaoaigaina talafeagai, e maua ai fa'amatalaga i luga o le cellular ma molecular process o lo'o fa'avaeina 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 o auaunaga.I le taimi lea, matou te ofoina atu le tulitatao o poloketi, fesoasoani faʻafitauli, ma Q&A sauniga e faʻatalanoa 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 |
| ua faaitiitia le rRNA | Illumina PE150 | 16 Gb | Q30≥85% |
| Tele ua filifilia | Illumina SE50 | 10-20M faitau |
Fa'ata'ita'iga Mana'oga:
Nucleotides:
| Conc.(ng/μl) | Aofa'i (μg) | mama | Fa'amaoni |
| ≥ 100 | ≥ 1 | 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. | La'au: RIN≥6.5 Manu: RIN≥7.0 5.0≥28S/18S≥1.0; fa'atapula'a pe leai se maualuga maualuga |
Fautuaina Fa'ata'ita'iga Tu'uina atu
pusa:
2 ml centrifuge paipa (E le fautuaina le pepa apa)
Fa'ailoga fa'ata'ita'iga: Vaega+fa'atusa eg A1, A2, A3;B1, B2, B3... ...
utaina:
1. Dry-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 faʻamago i le RNA stabilization tube(eg RNAstable®) ma lafo i le vevela potu.
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
Fa'amatalaga fa'amatalaga RNA
Genes Fa'aaliga Eseese
su'esu'ega ceRNA
MiRNA fa'aalia eseese ma RNA fa'atatau
Su'esu'e le alualu i luma o su'esu'ega na fa'afaigofieina e le BMKGene 'au'aunaga fa'asologa fa'asologa atoa e ala i le aoina o fa'asalalauga.
Dai, Y. et al.(2022) 'Faʻamatalaga faʻamatalaga lautele o mRNAs, lncRNAs ma miRNAs i le faʻamaʻi Kashin-Beck faʻamaonia e le RNA-sequencing', Molecular Omics, 18 (2), itulau 154-166.doi: 10.1039/D1MO00370D.
Liu, N. nan et al.(2022) 'Full length transcriptomes analysis of cold-resistance of Apis cerana in Changbai Mountain during overwintering period.', Gene, 830, itulau 146503–146503.doi: 10.1016/J.GENE.2022.146503.
Wang, XJ et al.(2022) 'Multi-Omics Integration-Based Priritization of Cometing Endogenous RNA Regulation Networks in Small Cell Lung Cancer: Molecular Characteristics and Drug Candidates', Frontiers in Oncology, 12, p.904865. doi: 10.3389/FONC.2022.904865/BIBTEX.
Xu, P. et al.(2022) 'Su'esu'ega tu'ufa'atasi o fa'amatalaga fa'amatalaga lncRNA/circRNA-miRNA-mRNA e fa'aalia ai ni fa'amatalaga fou i auala e mafai ona tali atu i nematodes a'a i le pinati', BMC Genomics, 23(1), itulau 1-12.doi: 10.1186/S12864-022-08470-3/ATA/7.
Yan, Z. et al.(2022) 'O le fa'asologa atoa o le RNA o lo'o fa'amamafaina ai le mole mole e feso'ota'i ma le tausiga o le lelei o le seleselega i le broccoli e ala i le fa'avelaina o le LED mumu', Postharvest Biology and Technology, 188, i.111878. doi: 10.1016/J.POSTHARVBIO.2022.111878.
