● Zvakanyanya Kuitika: Pamusoro pe200,000 samples dzakagadziriswa muBMK inovhara marudzi akasiyana-siyana emuenzaniso, kusanganisira sero tsika, tishu, mvura yemuviri, nezvimwewo uye pamusoro pe7,000 mRNA-Seq mapurojekiti akavharwa achivhara nzvimbo dzakasiyana-siyana dzekutsvakurudza.
● Strict quality control system: Core quality control points kuburikidza nematanho ese anosanganisira kugadzirira sampuli, kugadzirira raibhurari, sequencing uye bioinformatics zviri pasi pekutariswa kwepedyo kuitira kuti vape mhedzisiro yemhando yepamusoro.
● Madhatabheti akawanda anowanikwa echiziviso chebasa uye zvidzidzo zvekusimudzira kuzadzisa zvinangwa zvetsvagiridzo zvakasiyana.
● Mushure mekutengesa masevhisi: Mushure mekutengesa masevhisi anoshanda kwemwedzi mitatu pakupedzwa kweprojekiti, kusanganisira kutevera mapurojekiti, kupfura-dambudziko, mhinduro Q&A, nezvimwe.
Library | Sequencing strategy | Data inokurudzirwa | Quality Control |
Poly A yakafumiswa | Illumina PE150 | 6 Gb | Q30≥85% |
Nucleotides:
Conc.(ng/μl) | Mari (μg) | Kuchena | Kuperera |
≥ 20 | ≥ 0.5 | OD260/280=1.7-2.5 OD260/230=0.5-2.5 Yakaganhurirwa kana isina mapuroteni kana kusvibiswa kweDNA kunoratidzwa pagel. | Kumiti: RIN≥6.5; Zvemhuka: RIN≥7.0; 5.0≥28S/18S≥1.0; kukwirira kwakadzika kana kusina kwekutanga |
Tissue: Huremu (yakaoma):≥1 g
*Kune tishu diki pane 5 mg, tinokurudzira kutumira flash yakaomeswa nechando (mumvura nitrogen) sampuli yetishu.
Kumiswa kwesero:Nhamba yemasero = 3 × 1061 × 107
* Isu tinokurudzira kutumira chando cell lysate.Kana iyo sero inoverenga idiki pane 5 × 105.
Mienzaniso yeropa:Vhoriyamu≥1 ml
Microorganism:Husa ≥ 1 g
Container: 2 ml centrifuge chubhu (Tin foil haina kukurudzirwa)
Sample labeling: Boka+ replicate eg A1, A2, A3;B1, B2, B3...
Shipment:
Bioinformatics
Eukaryotic mRNA sequencing yekuongorora mafambiro
Bioinformatics
ØRaw data quality control
ØReference genome alignment
ØKuongororwa kwechimiro chezvinyorwa
ØExpression quantification
ØDifferential expression analysis
ØBasa rekuzivisa uye kusimudzira
1.mRNA Data Saturation curve
2.Differential expression analysis-Volcano plot
3.KEGG chirevo paDEGs
4.GO classification paDEGs