Me RNA sequencing-Illumina
Nta
● Kev xaiv loj ntawm RNA ua ntej kev npaj tsev qiv ntawv
● Bioinformatic tsom xam nyob ib ncig ntawm miRNA kev kwv yees thiab lawv lub hom phiaj
Kev Pabcuam Zoo
●Comprehensive bioinformatics tsom xam:Ua kom muaj kev txheeb xyuas ntawm ob qho tib si paub thiab tshiab miRNAs, kev txheeb xyuas ntawm miRNAs lub hom phiaj thiab cov lus qhia ua haujlwm tau zoo thiab txhawb nqa nrog ntau cov ntaub ntawv (KEGG, GO)
●Rigorous Quality Control: peb siv cov ntsiab lus tseem ceeb hauv txhua theem, los ntawm cov qauv thiab kev npaj tsev qiv ntawv mus rau kev sib txuas lus thiab bioinformatics.Qhov kev soj ntsuam nruj me ntsis ua kom cov khoom xa tuaj raws li cov txiaj ntsig zoo.
●Kev them nyiaj yug tom qab muag: Peb qhov kev cog lus txuas ntxiv dhau qhov kev ua tiav nrog rau 3 lub hlis tom qab kev muag khoom.Nyob rau lub sijhawm no, peb muab cov phiaj xwm taug qab, kev pab daws teeb meem, thiab Q&A ntu los daws cov lus nug ntsig txog cov txiaj ntsig.
●Kev txawj ntse: nrog cov ntaub ntawv teev tseg ntawm kev ua tiav kaw dhau ntau qhov haujlwm sRNA npog ntau dua 100 hom hauv ntau qhov kev tshawb fawb, peb pab pawg coj ntau yam kev paub rau txhua qhov haujlwm.
Sample Requirements thiab Delivery
| Lub tsev qiv ntawv | Platform | Cov ntaub ntawv pom zoo | Cov ntaub ntawv QC |
| Xaiv qhov loj me | Illumina SE 50 | 10M-20M nyeem | Q30≥85% |
Cov Qauv Xav Tau:
Nucleotides:
| Conc.(ng/μl) | Tus nqi (μg) | Purity | Kev ncaj ncees |
| ≥80 | ≥ 0.5 | OD 260/280 = 1.7-2.5 OD 260/230 = 0.5-2.5 Txwv tsis pub lossis tsis muaj protein ntau lossis DNA kis tau pom ntawm gel. | RIN ≥6.5; 5.0 ≥28S / 18S≥1.0; txwv los yog tsis muaj qhov nce siab |
● Nroj tsuag:
Hauv paus, qia los yog Petal: 450 mg
Nplooj los yog noob: 300 mg
Txiv Hmab Txiv Ntoo: 1.2 g
● Tsiaj:
Lub plawv lossis txoj hnyuv: 450 mg
Viscera lossis hlwb: 240 mg
Cov leeg: 600 mg
Pob txha, plaub hau lossis tawv nqaij: 1.5g
● Arthropods:
piv: 9g
Crustacea: 450 mg
● Tag nrho cov ntshav:2 cev
● Cells: 106 hlwb
● Ntshav thiab Plasma:6ml ib
Pom zoo coj mus kuaj
Ntim:
2 ml centrifuge raj (Tin foil tsis pom zoo)
Sample labeling: Pab pawg + replicate xws li A1, A2, A3;B1, B2, B3....
Kev xa khoom:
1.Dry-ice: Cov qauv yuav tsum tau ntim rau hauv hnab thiab muab faus rau hauv cov dej khov qhuav.
2.RNAstable raj: RNA cov qauv tuaj yeem qhuav hauv RNA stabilization raj (xws li RNAstable®) thiab xa mus rau hauv chav tsev kub.
Kev Pabcuam Ua Haujlwm Flow
Kev sim tsim
Qauv xa tuaj
RNA rho tawm
Kev tsim kho tsev qiv ntawv
Sequencing
Cov ntaub ntawv tsom xam
Kev pab cuam tom qab muag
Bioinformatics
Kev txheeb xyuas ntawm miRNA: qauv thiab qhov tob
Kev qhia txawv ntawm miRNA - hierarchical pawg
Cov lus piav qhia ua haujlwm ntawm lub hom phiaj ntawm qhov sib txawv qhia miRNAs
Tshawb nrhiav cov kev tshawb fawb nce qib uas tau tsim los ntawm BMKGene 'sRNA sequencing cov kev pabcuam los ntawm kev sau cov ntawv tshaj tawm.
Chen, H. et al.(2023) 'Viral infections inhibit saponin biosynthesis and photosynthesis in Panax notoginseng', Plant Physiology and Biochemistry, 203, p.108038. doi: 10.1016/J.PLAPHY.2023.108038.
Li, H. et al.(2023) ' Cov nroj tsuag FYVE domain-muaj protein ntau FREE1 koom nrog microprocessor Cheebtsam rau repress miRNA biogenesis ', EMBO qhia, 24(1).doi: 10.15252/EMBR.202255037/SUPPL_FILE/EMBR202255037-SUP-0004-SDATAFIG4.TIF.
Yus, J. et al.(2023) 'Lub MicroRNA Ame-Bantam-3p Tswj Larval Pupal Kev Txhim Kho los ntawm Lub Hom Phiaj Ntau Yam Kev Loj Hlob Zoo li Domains 8 Gene (megf8) hauv Honeybee, Apis mellifera', International Journal of Molecular Sciences, 24(6), p .5726. doi: 10.3390/IJMS24065726/S1.
Zhang, M. Y. et al.(2018) 'Integrated Analysis of MiRNA and Genes Associated with Meat Quality Reveals that Gga-MiR-140-5p Affects Intramuscular Fat Deposition in Chickens', Cellular Physiology and Biochemistry, 46(6), pp. 2421–2433.doi: 10.1159/000489649 ib.
