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Eukaryotic mRNA sequencing-Illumina

mRNA sequencing inogonesa profiling yeese mRNAs akanyorwa kubva kumaseru pasi pemamiriro chaiwo.Iyo tekinoroji ine simba yekuburitsa chimiro chejeni, zvimiro zvemajini uye mamorekuru emamwe maitiro ebhayoloji.Parizvino, kutevedzana kwemRNA kwave kushandiswa zvakanyanya mukutsvaga kwakakosha, kuongororwa kwekiriniki, kukura kwezvinodhaka, nezvimwe.

Platform: Illumina NovaSeq 6000


Service Details

Bioinformatics

Demo Results

Zvakanakira

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.

Sample Zvinodiwa uye Delivery

Library Sequencing strategy Data inokurudzirwa Quality Control
Poly A yakafumiswa Illumina PE150

6 Gb

Q30≥85%

Sample Zvinodiwa:

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

Yakakurudzirwa Sample Delivery

Container: 2 ml centrifuge chubhu (Tin foil haina kukurudzirwa)

Sample labeling: Boka+ replicate eg A1, A2, A3;B1, B2, B3...

Shipment:

  1. Dry-ice: Samples dzinofanirwa kutakurwa mumabhegi uye kuvigwa mune yakaoma-aizi.
  2. RNAstable chubhu: RNA samples inogona kuomeswa muRNA kudzikamisa chubhu (eg RNAstable®) uye kutumirwa mukamuri tembiricha.

Kuyerera Kwebasa Rebasa

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Kuedza kugadzira

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Muenzaniso wekutumira

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RNA kubviswa

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Kuvakwa kweraibhurari

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Sequencing

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Data analysis

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Mushure mekutengesa masevhisi


  • Zvakapfuura:
  • Zvinotevera:

  • Bioinformatics

    2(1)

    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

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    2.Differential expression analysis-Volcano plot

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    3.KEGG chirevo paDEGs

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    4.GO classification paDEGs

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