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Cov khoom

Eukaryotic mRNA sequencing-Illumina

mRNA sequencing enables profiling ntawm tag nrho cov mRNAs transcripted los ntawm hlwb nyob rau hauv tej yam kev mob.Nws yog lub tshuab muaj zog rau kev nthuav tawm cov noob qhia profile, cov qauv noob thiab cov txheej txheem molecular ntawm qee cov txheej txheem lom neeg.Txog niaj hnub no, mRNA sequencing tau dav siv hauv kev tshawb fawb, kev kuaj mob, kev tsim tshuaj, thiab lwm yam.

Platform: Illumina NovaSeq 6000


Kev Paub meej

Bioinformatics

Cov txiaj ntsig Demo

Qhov zoo

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.

Sample Requirements thiab Delivery

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%

Cov Qauv Xav Tau:

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

Pom zoo coj mus kuaj

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:

  1. Qhuav-dej: Cov qauv yuav tsum tau ntim rau hauv hnab thiab faus rau hauv cov dej khov qhuav.
  2. RNAstable raj: RNA cov qauv tuaj yeem qhuav hauv RNA stabilization raj (xws li RNAstable®) thiab xa mus rau hauv chav tsev kub.

Kev Pabcuam Ua Haujlwm Flow

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Kev sim tsim

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Qauv xa tuaj

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RNA rho tawm

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Kev tsim kho tsev qiv ntawv

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Sequence

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Cov ntaub ntawv tsom xam

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Kev pab cuam tom qab muag


  • Yav dhau los:
  • Tom ntej:

  • Bioinformatics

    2(1)

    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

    3(1)

    2.Differential expression analysis-Volcano plot

    4(1)

    3.KEGG annotation rau DEGs

    5(1)

    4.GO kev faib tawm ntawm DEGs

    6(1)

    tau ib nqe lus

    Sau koj cov lus ntawm no thiab xa tuaj rau peb

    Xa koj cov lus rau peb: