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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 Platform


Service Details

Bioinformatics

Demo Results

Zvakanakira

● 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.

Sample Zvinodiwa uye Delivery

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

6 Gb

Q30≥85%

Sample Zvinodiwa:

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

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

Muenzaniso QC

Kuedza kugadzira

sampuli yekuendesa

Muenzaniso wekutumira

Pilot experiment

RNA kubviswa

Kugadzirira Raibhurari

Kuvakwa kweraibhurari

Sequencing

Sequencing

Data analysis

Data analysis

Mushure mekutengesa Services

Mushure mekutengesa masevhisi


  • Zvakapfuura:
  • Zvinotevera:

  • Bioinformatics

    wps_doc_10

    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

    3(1)

    2.Differential expression analysis-Volcano plot

    4(1)

    3.KEGG chirevo paDEGs

    5(1)

    4.GO classification paDEGs

    6(1)

    kuwana quote

    Nyora meseji yako pano uye titumire kwatiri

    Tumira meseji yako kwatiri: