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Zogulitsa

Eukaryotic mRNA sequencing-Illumina

Kutsatizana kwa mRNA kumathandizira kuti mbiri ya mRNA yonse yolembedwa kuchokera ku cell ikhale yodziwika.Ndi ukadaulo wamphamvu wowulula mbiri ya jini, kapangidwe ka majini ndi mamolekyu azinthu zina zachilengedwe.Mpaka pano, kutsatizana kwa mRNA kwagwiritsidwa ntchito kwambiri pakufufuza kofunikira, zowunikira zamankhwala, chitukuko chamankhwala, ndi zina zambiri.

Platform: Illumina NovaSeq 6000


Tsatanetsatane wa Utumiki

Bioinformatics

Zotsatira za Demo

Ubwino wake

ØOdziwika Kwambiri: Zitsanzo zopitilira 200,000 zasinthidwa mu BMK zomwe zikukhudza mitundu yosiyanasiyana, kuphatikiza chikhalidwe cha maselo, minofu, madzi amthupi, ndi zina zambiri.

ØDongosolo lokhazikika loyang'anira khalidwe: Malo owongolera khalidwe lapakati pamasitepe onse kuphatikizapo kukonzekera zitsanzo, kukonzekera laibulale, kutsatizana ndi bioinformatics akuyang'aniridwa kuti apereke zotsatira zapamwamba kwambiri.

ØMa database angapo omwe amapezeka kuti afotokoze ntchito ndi maphunziro owonjezera kuti akwaniritse zolinga zosiyanasiyana za kafukufuku.

ØNtchito zogulitsa pambuyo pogulitsa: Ntchito zogulitsa pambuyo pake ndizovomerezeka kwa miyezi itatu polojekiti ikamalizidwa, kuphatikiza kutsata ma projekiti, kuwombera zovuta, zotsatira za Q&A, ndi zina.

Zitsanzo Zofunika ndi Kutumiza

Library Njira yotsatirira Deta yovomerezeka Kuwongolera Kwabwino
Poly A yowonjezera Chithunzi cha PE150

6gb pa

Q30≥85%

Zitsanzo Zofunika:

Nucleotides:

Chiyero Umphumphu Ndalama
OD260/280≥1.7-2.5 OD260/230≥0.5-2.5Zochepa kapena zopanda mapuloteni kapena DNA kuipitsidwa komwe kumawonetsedwa pa gel. Zazomera: RIN≥6.5;Zanyama: RIN≥7;28S/18S≥1.0; kukwera kochepa kapena kulibe koyambira Conc.≥30 ng/μl;Kuchuluka ≥ 10 μl;Zonse ≥ 1.5 μg

Thupi: Kulemera (kuuma):≥1 g
*Pa minofu yaing'ono kuposa 5 mg, timalimbikitsa kutumiza zitsanzo za minofu yowuma (mu madzi a nayitrogeni).

Kuyimitsidwa kwa cell:Chiwerengero cha ma cell = 3 × 106-1 × 107
*Tikupangira kutumiza frozen cell lysate.Ngati cellyo ikhala yaying'ono kuposa 5 × 105, kung'anima kozizira mu nayitrogeni wamadzimadzi kumalimbikitsidwa, komwe kumakhala koyenera kutulutsa pang'ono.

Zitsanzo za magazi:Kuchuluka ≥1 ml

Microorganism:Kulemera ≥ 1 g

Kutumiza Kwachitsanzo Kovomerezeka

Chidebe: 2 ml centrifuge chubu (zojambula za malata ndizosavomerezeka)

Zitsanzo zolembera: Gulu+ lobwereza mwachitsanzo A1, A2, A3;B1, B2, B3...

Kutumiza:

  1. Dry-ice: Zitsanzo ziyenera kupakidwa m'matumba ndikukwiriridwa mu ayezi wouma.
  2. RNAstable machubu: RNA zitsanzo akhoza zouma mu RNA kukhazikika chubu (mwachitsanzo RNAstable®) ndi kutumizidwa mu firiji.

Kuyenda kwa Ntchito Yantchito

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Kuyesera kupanga

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Kutumiza kwachitsanzo

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Kusintha kwa RNA

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Kumanga laibulale

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Kutsata

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Kusanthula deta

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Pambuyo pogulitsa ntchito


  • Zam'mbuyo:
  • Ena:

  • Bioinformatics

    2(1)

    Eukaryotic mRNA sequencing kusanthula kayendedwe ka ntchito

    Bioinformatics

    ØKuwongolera khalidwe la data yaiwisi

    ØReference genome alignment

    ØKusanthula kapangidwe ka zilembo

    ØKuchulukitsa kwa mawu

    ØKusanthula kwamawu osiyanasiyana

    ØTanthauzo la ntchito ndi kuwonjezera

    1.mRNA Data Saturation curve

    3(1)

    2.Differential expression analysis-Volcano plot

    4(1)

    3.Ndemanga za KEGG pa DEGs

    5(1)

    4.GO classification pa DEGs

    6(1)

    pezani mtengo

    Lembani uthenga wanu apa ndikutumiza kwa ife

    Titumizireni uthenga wanu: