China Eukaryotic mRNA ngokulandelelana-Illumina umzi-mveliso kunye nabavelisi |BMK

Ulandelelwano lwe-Eukaryotic mRNA-Illumina

Eukaryotic mRNA sequencing-Illumina Featured Image

Ulandelelwano lwe-mRNA lwenza ukuba iprofayili yazo zonke i-mRNA zikhutshelwe kwiiseli phantsi kweemeko ezithile.Bubuchwephesha obunamandla bokutyhila inkangeleko yemfuza yemfuza, izakhiwo zemfuza kunye neendlela zemolekyuli zeenkqubo ezithile zebhayoloji.Ukuza kuthi ga ngoku, ulandelelwano lwe-mRNA lusetyenziswe ngokubanzi kuphando olusisiseko, uxilongo lwezonyango, uphuhliso lwamachiza, njl.

Iqonga: Illumina NovaSeq 6000

Qhagamshelana nathi

Iinkcukacha zeNkonzo

Bioinformatics

Iziphumo zeDemo

Iingenelo

ØAmava aphezulu: Zingaphezu kwama-200,000 iisampulu ezisetyenzisiweyo kwi-BMK egubungela iindidi ezahlukeneyo zeesampulu, kubandakanywa inkcubeko yeeseli, izicubu, ulwelo lomzimba, njl.njl. kunye neeprojekthi ezingaphezu kwe-7,000 ze-mRNA-Seq ezivaliweyo ezibandakanya iindawo ezahlukeneyo zophando.

ØInkqubo engqongqo yolawulo lomgangatho: Amanqaku olawulo lomgangatho angundoqo kuwo onke amanyathelo abandakanya ukulungiswa kwesampulu, ukulungiswa kwethala leencwadi, ulandelelwano kunye ne-bioinformatics ziphantsi kweliso elibukhali ukuze kunikezelwe iziphumo ezikumgangatho ophezulu.

ØIindawo ezininzi zogcino-lwazi ezifumanekayo zochazo lomsebenzi kunye nezifundo zokutyebisa ukuzalisekisa iinjongo zophando ezahlukeneyo.

ØIinkonzo zokuthengisa emva kokuthengiswa: Iinkonzo emva kokuthengiswa zivumelekile kwiinyanga ezi-3 ekugqityweni kweprojekthi, kubandakanywa ukulandelwa kweeprojekthi, ukuxazulula iingxaki, iziphumo ze-Q & A, njl.

IiMfuno zeSampulu kunye nokuhanjiswa

Ithala leencwadi	Isicwangciso sokulandelelana	Idatha iyacetyiswa	Ulawulo lwemeko
UPoly A utyebile	Illumina PE150	6Gb	Q30≥85%

IiMfuno zeSampulu:

IiNucleotides:

Ubunyulu	Ingqibelelo	Isixa
OD260/280≥1.7-2.5 OD260/230≥0.5-2.5Unyino okanye akukho protein okanye ungcoliseko lweDNA oluboniswa kwijeli.	Kwizityalo: RIN≥6.5;Izilwanyana: RIN≥7;28S/18S≥1.0;umphakamo oqingqiweyo okanye akukho siseko	Conc.≥30 ng/μl;Umthamo ≥ 10 μl;Iyonke ≥ 1.5 μg

Izicubu: Ubunzima (bomile):≥1 g
*Kwithishu ezincinci kuno-5 mg, sicebisa ukuba uthumele ifleshi ekhenkcezisiweyo (kwinitrogen engamanzi) isampulu yethishu.

Ukumiswa kweeseli:Ubalo lweeseli = 3×10⁶- 1×10⁷
*Sincoma ukuhambisa i-lysate yeseli enomkhenkce.Kwimeko apho iseli ibala encinci kune-5 × 105, i-flash ekhenkcezwe kwinitrogen engamanzi iyacetyiswa, ekhethwayo xa kutsalwa kancinci.

Iisampulu zegazi:Umthamo≥1 ml

I-Microorganism:Ubunzima ≥ 1 g

Unikezelo lweSampuli eCetyisiweyo

Isikhongozeli: 2 ml centrifuge ityhubhu (Ifoyile yeTin ayikhuthazwa)

Isampuli yokuleyibhela: Iqela+lokuphindaphinda umz. A1, A2, A3;B1, B2, B3... ...

Ukuthunyelwa ngenqanawa:

Umkhenkce owomileyo: Iisampulu kufuneka zipakishwe kwiingxowa kwaye zingcwatywe kwindawo eyomileyo.
Iityhubhu ze-RNAstable: Iisampulu ze-RNA zingomiswa kwityhubhu yokuzinzisa ye-RNA (umzekelo, i-RNAstable®) kwaye ithunyelwe kwindawo yobushushu begumbi.