ØZvakanyanya Kuitika: Pamusoro pe200,000 samples dzakagadziriswa muBMK ichivhara akasiyana emhando dzemuenzaniso, kusanganisira sero tsika, tishu, mvura yemuviri, nezvimwewo uye anopfuura zviuru zvinomwe zveMRNA-Seq mapurojekiti akavharwa achivhara nzvimbo dzakasiyana dzekutsvagisa.
ØStrict quality control system: Core quality control points kuburikidza nematanho ese anosanganisira kugadzirira sampuli, kugadzirira raibhurari, kutevedzana uye bioinformatics zviri pasi pekutariswa kwepedyo kuitira kuti vape mhedzisiro yemhando yepamusoro.
ØAkawanda dhatabhesi anowanikwa ekuita zvirevo uye zvidzidzo zvekusimudzira kuzadzisa zvinangwa zvetsvagiridzo zvakasiyana.
ØMushure 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:
Kuchena | Kuperera | Mari |
OD260/280≥1.7-2.5 OD260/230≥0.5-2.5Yakaganhurirwa kana isina mapuroteni kana kusvibiswa kweDNA kunoratidzwa pagel. | Zvezvirimwa: RIN≥6.5;Zvemhuka: RIN≥7;28S/18S≥1.0;inogumira kana isina kukwirira kwekutanga | Conc.≥30 ng/μl;Vhoriyamu ≥ 10 μl;Zvose ≥ 1.5 μg |
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 ichiverengera idiki pane 5 × 105, flash yakaomeswa mune yemvura nitrogen inokurudzirwa, iyo inosarudzika kudhirowa diki.
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