ØKev Ua Tau Zoo: Ntau tshaj 200,000 tus qauv tau ua tiav hauv BMK npog ntau hom qauv, suav nrog kab lis kev cai ntawm tes, ntaub so ntswg, kua dej hauv lub cev, thiab lwm yam thiab tshaj li 7,000 mRNA-Seq tej yaam num kaw npog ntau yam kev tshawb fawb cheeb tsam.
ØKev Tswjhwm nruj nruj: Cov ntsiab lus tseem ceeb ntawm kev tswj xyuas zoo dhau los ntawm txhua kauj ruam suav nrog kev npaj ua qauv, npaj tsev qiv ntawv, ua ntu zus thiab bioinformatics yog nyob rau hauv kev saib xyuas kom ua tau zoo.
ØNtau cov ntaub ntawv muaj rau kev ua haujlwm annotation thiab kev tshawb fawb ntxiv kom ua tiav ntau hom kev tshawb fawb.
ØCov kev pab cuam tom qab muag: Cov kev pab cuam tom qab muag siv tau rau 3 lub hlis tom qab ua tiav qhov project, suav nrog cov haujlwm ua raws, teeb meem-tua, cov txiaj ntsig Q&A, thiab lwm yam.
Lub tsev qiv ntawv | Sequencing tswv yim | Cov ntaub ntawv pom zoo | Kev Tswj Xyuas Zoo |
Poly A enriched | Illumina PE 150 | 6gb ib | Q30≥85% |
Nucleotides:
Purity | Kev ncaj ncees | Tus nqi |
OD260/280≥1.7-2.5 OD260/230≥0.5-2.5Txwv los yog tsis muaj protein ntau lossis DNA kis tau pom ntawm gel. | Rau cov nroj tsuag: RIN ≥6.5; Rau tsiaj: RIN≥7; 28S / 18S≥1.0; txwv lossis tsis muaj qhov siab | Conc.≥30 ng / μl;Ntim ≥ 10 μl;Tag nrho ≥ 1.5 μg |
Cov ntaub so ntswg: Nyhav (qhuav):≥1g
* Rau cov ntaub so ntswg me dua 5 mg, peb pom zoo kom xa flash khov (hauv kua nitrogen) cov ntaub so ntswg.
Cell ncua:Cell suav = 3 × 106- 1 × 107
* Peb pom zoo kom nkoj khov cell lysate.Nyob rau hauv rooj plaub uas cov xov tooj suav me dua 5 × 105, flash khov hauv cov kua nitrogen yog pom zoo, uas yog qhov zoo dua rau kev rho tawm micro.
Cov ntshav kuaj:Ntim ≥ 1 ml
Microorganism:Qhov hnyav ≥ 1 g
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:
Bioinformatics
Eukaryotic mRNA sequencing tsom xam workflow
Bioinformatics
ØRaw cov ntaub ntawv zoo tswj
ØReference genome alignment
ØTranscript structure analysis
ØKev nthuav qhia kom muaj nuj nqis
ØDifferential expression analysis
ØFunction annotation thiab enrichment
1.mRNA Data Saturation nkhaus
2.Differential expression analysis-Volcano plot
3.KEGG annotation rau DEGs
4.GO kev faib tawm ntawm DEGs