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Matsakaicin tushen mRNA Sequencing-Illumina

mRNA sequencing ya rungumi dabara na gaba-gaba (NGS) don kama manzo RNA (mRNA) ya samar da Eukaryote a takamaiman lokacin da wasu ayyuka na musamman ke kunnawa.Mafi tsayin rubutun da aka raba ana kiransa 'Unigene' kuma an yi amfani da shi azaman jeri na bincike na gaba, wanda shine ingantacciyar hanya don nazarin tsarin kwayoyin halitta da hanyar sadarwa na tsarin jinsin ba tare da tunani ba.

Bayan tattara bayanan kwafi da bayanin aikin unigene

(1) Binciken SSR, tsinkayar CDS da tsarin kwayoyin halitta za a riga an tsara su.

(2) Ƙididdigar maganganun unigene a cikin kowane samfurin za a yi.

(3)Unigenes daban-daban da aka bayyana tsakanin samfurori (ko ƙungiyoyi) za a gano su bisa lafazin unigene

(4) Za a gudanar da tari, bayanin aiki da kuma ingantaccen bincike na unigenes da aka bayyana daban-daban.


Cikakkun Sabis

Bioinformatics

Sakamakon Demo

Nazarin Harka

Siffofin

● Ba tare da kowane nau'in kwayar halitta ba,

Za'a iya amfani da bayanan don nazarin tsari da kuma bayanin rubutun

● Gano madaidaitan wuraren yanka

Amfanin Sabis

● Bayar da sakamako na tushen BMKCloud: Ana ba da sakamakon azaman fayil ɗin bayanai da rahoton hulɗa ta hanyar dandamali na BMKCloud, wanda ke ba da damar karantawa mai amfani mai amfani na abubuwan bincike mai rikitarwa da ma'adinan bayanan da aka keɓance akan tushen daidaitaccen bincike na bioinformatics.

● Bayan-sayar da sabis: Bayan-sayar da sabis na aiki na tsawon watanni 3 bayan kammala aikin, gami da bin diddigin ayyukan, harbin matsala, Q&A sakamakon, da sauransu.

Samfuran Bukatun da Bayarwa

Samfuran Bukatun:

Nucleotides:

Conc.(ng/μl)

Adadin (μg)

Tsafta

Mutunci

≥ 20

≥ 0.5

OD260/280=1.7-2.5

OD260/230=0.5-2.5

Iyakance ko babu furotin ko gurɓataccen DNA da aka nuna akan gel.

Don tsire-tsire: RIN≥6.5;

Na dabbobi: RIN≥7.0;

5.0≥28S/18S≥1.0;

iyakance ko babu hawan tushe

Nama: Nauyi (bushe): ≥1 g
* Don nama ƙasa da 5 MG, muna ba da shawarar aika samfurin nama daskararre (a cikin ruwa nitrogen).

Dakatar da salula: Ƙididdiga ta salula = 3×107
*Muna ba da shawarar jigilar daskararrun cell lysate.Idan wannan tantanin halitta ya ƙidaya ƙasa da 5 × 105, walƙiya daskararre a cikin ruwa nitrogen ana shawarar.

Samfuran jini:
PA×geneBloodRNATube;
6mLTRIzol da 2ml jini (TRIzol:Blood=3:1)

Isar da Samfurin Nasiha

Kwantena:
2 ml bututu centrifuge (Ba a ba da shawarar foil tin ba)
Alamar samfur: Ƙungiya+ kwafi misali A1, A2, A3;B1, B2, B3......

Jirgin ruwa:
1.Dry-ice: Ana buƙatar samfurori a cikin jaka kuma a binne su a bushe-kankara.
2.RNAstable tubes: Ana iya bushe samfuran RNA a cikin bututun daidaitawar RNA (misali RNAstable®) kuma a tura su cikin zafin jiki.

Gudun Aikin Sabis

Misalin QC

Gwajin ƙira

samfurin bayarwa

Samfurin bayarwa

Gwajin matukin jirgi

RNA cirewa

Shirye-shiryen Laburare

Gina ɗakin karatu

Jeri

Jeri

Binciken bayanai

Binciken bayanai

Bayan Sabis na siyarwa

Bayan-sayar da sabis


  • Na baya:
  • Na gaba:

  • Bioinformatics

    wps_doc_11

    1.mRNA(denovo) Ka'idar Taro

    Ta Triniti, an raba karatun zuwa ƙananan ƙananan, wanda aka sani da K-mer.Ana amfani da waɗannan K-mers azaman tsaba da za'a faɗaɗa su zuwa ƙugiya sannan kuma ana dogara da abubuwan da ke kan juzu'i.A ƙarshe, an yi amfani da De Bruijn a nan don gane kwafi a cikin abubuwan.

    mRNA-(De-novo)-Bayyana-na-Triniti

    mRNA (De novo) Bayanin Triniti

    2.mRNA (De novo) Rarraba Matsayin Maganar Halitta

    RNA-Seq yana da ikon cimma ƙima mai mahimmanci na maganganun kwayoyin halitta.A al'ada, iya gano kewayon bayanan kwafi FPKM yana jere daga 10^-2 zuwa 10^6.

    mRNA-(De-novo)-Rarraba-na-FPKM-yawan-a cikin-kowane-samfurin

    mRNA (De novo) Rarraba yawan FPKM a cikin kowane samfurin

    3.mRNA (De novo) GO Analysis Enrichment Analysis of DEGs

    GO (Gene Ontology) ma'ajin bayanai shine ingantaccen tsarin bayanin halitta wanda ke ƙunshe da daidaitattun ƙamus na ayyukan samfuran kwayoyin halitta.Ya ƙunshi matakan da yawa, inda ƙananan matakin yake, mafi ƙayyadaddun ayyuka.

    mRNA-(De-novo) - GO-rarrabuwa-na-DEGs-a-mataki-biyu

    mRNA (De novo) GO rarrabuwa na DEGs a matakin na biyu

    BMK Case

    Nazari na Rubuce-rubuce na Sucrose Metabolism a Lokacin Bunƙasa Bulb da Ci gaban Albasa (Allium cepa L.)

    Buga: iyakoki a kimiyyar shuka,2016

    Dabarun jeri

    Illumina HiSeq2500

    Tarin samfurin

    Utah Yellow Sweet Spain cultivar "Y1351" an yi amfani da ita a cikin wannan binciken.Yawan samfuran da aka tattara shine
    15th Day bayan kumburi (DAS) na kwan fitila (2-cm diamita da 3-4 g nauyi), 30th DAS (5-cm diamita da 100-110 g nauyi), da ~ 3 a kan 40th DAS (7-cm diamita da 260-300 g).

    Sakamako mai mahimmanci

    1. a cikin zane na Venn, an gano jimlar 146 DEGs a duk nau'i-nau'i uku na matakan ci gaba.
    2. "Carbohydrate Transport and metabolism" an wakilta ta kawai 585 unigenes (watau 7% na COG annotated).
    3.Unigenes da aka samu nasarar bayyanawa ga GO database an kasaftasu zuwa manyan sassa uku don matakai daban-daban guda uku na ci gaban kwan fitila.Yawancin waɗanda aka wakilta a cikin “tsarin nazarin halittu” babban nau'in sune "tsarin ƙwayar cuta", sannan "tsarin salula".A cikin babban nau'in "ayyukan kwayoyin halitta" nau'ikan biyu da aka fi wakilta sune "dauri" da "aiki mai kara kuzari".

    PB-cikakken-tsawon-RNA-Sequencing-case-binciken

    Histogram na gungu na ƙungiyoyin orthologous (COG).

    PB-cikakken-tsawon-RNA-Sequencing-case-binciken

    Histogram na Gene Ontology (GO) rarrabuwa don unigenes da aka samo daga kwararan fitila a cikin matakai uku na haɓakawa.

    PB-cikakken-tsawon-RNA-Sequencing-case-binciken

    Zane na Venn yana nuna kwayoyin halitta daban-daban da aka bayyana a kowane matakai biyu na haɓaka kwan fitilar albasa

    Magana

    Zhang C, Zhang H, Zhan Z, da dai sauransu.Fassarar Fassarar Tattalin Arziki na Sucrose Metabolism a Lokacin Bunbulb da Ci Gaba a Albasa (Allium cepa L.) [J].Iyaka a Kimiyyar Shuka, 2016, 7:1425-.DOI: 10.3389/fpls.2016.01425

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