Laititi RNA Sequencing-Illumina
Vaega
● O sauniuniga a le faletusi e aofia ai se laasaga e filifili ai le lapopoa
● Bioinformatic su'esu'ega e fa'atotonugalemu ile va'aiga o le miRNA ma a latou fa'amoemoe
Tulaga Auaunaga
●Au'ili'iliga bioinformatics atoatoa:Fa'atagaina le fa'ailoaina o miRNAs e ta'uta'ua ma tala fou, fa'ailoaina o fa'amoemoega miRNA, ma fa'amatalaga fa'atino ma fa'atamaoaigaina i le tele o fa'amaumauga (KEGG, GO)
●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.
●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 faʻafeiloaʻi ai soʻo se fesili e fesoʻotaʻi ma iʻuga.
●Tomai Tele: Faatasi ai ma se faamaumauga o le tapunia manuia o le tele o poloketi sRNA e aofia ai le silia ma le 300 ituaiga i vaega eseese o suʻesuʻega, o la matou 'au e aumaia le tamaoaiga o le poto masani i galuega uma.
Fa'ata'ita'iga Mana'oga ma le Tu'uina atu
| Faletusi | Fa'avae | Fa'amatalaga fautuaina | Fa'amatalaga QC |
| Tele ua filifilia | Illumina SE50 | 10M-20M faitau | Q30≥85% |
Fa'ata'ita'iga Mana'oga:
Nucleotides:
| Conc.(ng/μl) | Aofa'i (μg) | Mama | Fa'amaoni |
| ≥ 80 | ≥ 0.8 | 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. | RIN≥6.0; 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:
Loto po'o le Intestinal: 450 mg
Viscera po'o Fai'ai: 240 mg
Musu: 600 mg
Ivi, Laulu po'o Pa'u: 1.5g
● Arthropods:
Iniseti: 9g
Crustacea: 450 mg
● toto atoa: 2 paipa
● Selula: 106 sela
● Serum ma Plasma:6 mL
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
● Pulea lelei o fa'amaumauga mata'utia
● fa'avasegaga sRNA
● Fa'atatau i se genome fa'asinomaga
● Fa'ailoaina ole miRNA ua iloa ma fou
● Su'esu'ega fa'amatalaga miRNA eseese
● Fa'amatalaga fa'atino o sini miRNA
Faailoaina o le miRNA: fausaga ma le loloto
Fa'aaliga 'ese'ese o le miRNA - fa'apipi'i fa'apitoa
Fa'amatalaga fa'atino o le fa'amoemoe o miRNA fa'aalia fa'aalia
Su'esu'e le alualu i luma o su'esu'ega na fa'afaigofieina e le BMKGene' sRNA sequencing services e ala i le aoina o fa'asalalauga.
Chen, H. et al. (2023) 'O fa'ama'i fa'ama'i e taofia ai le biosynthesis saponin ma le photosynthesis i Panax notoginseng', Plant Physiology and Biochemistry, 203, i. 108038. doi: 10.1016/J.PLAPHY.2023.108038.
Li, H. et al. (2023) 'O le laau FYVE domain-containing protein FREE1 e fesoʻotaʻi ma vaega microprocessor e taofiofia le miRNA biogenesis', EMBO lipoti, 24(1). doi: 10.15252/EMBR.202255037/SUPPL_FILE/EMBR202255037-SUP-0004-SDATAFIG4.TIF.
Yu, J. et al. (2023) 'O le MicroRNA Ame-Bantam-3p Pulea le Atina'eina o Pupal Larval e ala i le Fa'aautuina o le Fa'atupuina Fa'atupu Fa'atupu Fa'a-pei o Domains 8 Gene (megf8) i le Honeybee, Apis mellifera', International Journal of Molecular Sciences, 24(6), p . 5726. doi: 10.3390/IJMS24065726/S1.
Zhang, M. et al. (2018) 'Suʻesuʻega Tuʻufaʻatasia o MiRNA ma Genes e fesoʻotaʻi ma Aano o manufasi e faʻaalia ai o le Gga-MiR-140-5p e aʻafia ai le faʻaogaina o gaʻo i totonu o moa', Cellular Physiology and Biochemistry, 46 (6), itulau 2421-2433. doi: 10.1159/000489649.
