Ntev Tsis-coding Sequencing-Illumina
Kev Pabcuam Zoo
●Kev sib koom ua ke ntawm mRNA thiab lncRNA: los ntawm kev sib txuas cov lej ntawm mRNA cov ntawv sau tseg nrog kev kawm ntawm lncRNA thiab lawv lub hom phiaj, nws muaj peev xwm kom tau txais qhov tob tob ntawm cov txheej txheem tswj hwm hauv qab ntawm cov lus teb ntawm tes.
●Kev txawj ntse: Peb pab neeg nqa cov kev paub dhau los rau txhua qhov haujlwm, nrog rau cov ntaub ntawv teev tseg ntawm kev ua tiav ntau dua 23,000 tus qauv ntawm BMK hla ntau hom qauv thiab lncRNA cov haujlwm.
●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 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.
Sample Requirements thiab Delivery
| Lub tsev qiv ntawv | Platform | Cov ntaub ntawv pom zoo | Cov ntaub ntawv QC |
| rRNA depleted directional tsev qiv ntawv | Illumina PE 150 | 10-16 Gb | Q30≥85% |
Nucleotides:
| Conc.(ng/μl) | Tus nqi (μg) | Purity | Kev ncaj ncees |
| ≥80 | ≥ 0.8 | OD 260/280 = 1.7-2.5 OD 260/230 = 0.5-2.5 Txwv los yog tsis muaj protein ntau lossis DNA kis tau pom ntawm gel. | RIN ≥6.0; 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
● Cov ntshav tag nrho:2 cev
● Cells: 106 hlwb
● Ntshav thiab Plasmaib: 6ml
Pom zoo coj mus kuaj
Ntim: 2 ml centrifuge tube (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 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
Differential Gene Expression (DEGs) tsom xam
Quantification of lncRNA expression – clustering
Enrichment ntawm lncRNA lub hom phiaj noob
Kev sib koom ua ke mRNA thiab lncRNA txoj haujlwm tsom xam - Circos zajlus (lub voj voog nruab nrab yog mRNA thiab sab hauv cicrlce yog lncRNA)
Tshawb nrhiav cov kev nce qib uas tau pab txhawb los ntawm BMKGene's lncRNA equencing cov kev pabcuam los ntawm kev sau cov ntawv tshaj tawm.
Ji, H. et al. (2020) 'Kev txheeb xyuas, kev kwv yees ua haujlwm, thiab qhov tseem ceeb lncRNA kev txheeb xyuas qhov kev ntxhov siab ntsig txog lncRNAs hauv nas lub siab', Kev Tshaj Tawm Txog Kev Tshawb Fawb 2020 10: 1, 10(1), pp. 1–14. doi: 10.1038/s41598-020-57451-7 ib.
Jia, Z. et al. (2021) 'Integrative Transcriptomic Analysis Reveals the Immune Mechanism for a CyHV-3-Resistant Common Carp Strain', Frontiers in Immunology, 12, p. 687151. doi: 10.3389/FIMMU.2021.687151/BIBTEX.
Wang, XJ et al. (2022) 'Multi-Omics Integration-Based Prioritization of Competing 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.
Xiao, L. et al. (2020) 'Cov noob caj ces ntawm cov noob caj noob ces coexpression network hauv qab photosynthesis hauv Populus', Plant Biotechnology Journal, 18(4), pp. 1015–1026. doi: 10.1111/PBI.13270 ib.
Zheng H., et al. (2022) 'Lub Ntiaj Teb Kev Tswj Xyuas Kev Ruaj Ntseg rau Dysregulated Gene Expression thiab Abnormal Metabolic Signaling in Immune Cells in Microenvironment of Graves' Disease and Hashimoto's Thyroiditis', Frontiers in Immunology, 13, p. 879824. doi: 10.3389/FIMMU.2022.879824/BIBTEX.
