China Eukaryotic mRNA sequencing-Illumina fakitale ndi opanga |BMK

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

Lumikizanani nafe

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:

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