V8 RNA-seq Library Prep Kit ya RNA Library Yomanga: Pezani Mawu, Kufunsa, Mtengo Kapena Gulani

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VAHTS Universal V8 RNA-seq Library Prep Kit ya Illumina imagwiritsidwa ntchito pokonzekera malaibulale a transcriptome a nsanja za Illumina.Chidacho ndi chosunthika komanso choyenera pakupanga laibulale ya RNA ya RNA yopezedwa ndi Poly(A)-based mRNA enrichment kapena rRNA depletion.
Chidachi chili ndi mitundu iwiri ya cDNA 2nd Strand synthesis buffers. Mabuffers amatha kusankhidwa kuti amange malaibulale kuti athe kusanthula zolembalemba za RNA-Seq zosasinthika kapena zosasinthika. ntchito yomanga laibulale ndikuchepetsa nthawi yogwira ntchito.
The wokometsedwa anachita dongosolo bwino laibulale ntchito yomanga, amapereka yunifolomu Kuphunzira kwa ndalama zosiyanasiyana athandizira RNA, ndipo n'zogwirizana ndi otsika athandizira RNAs.Atasankha kukula ntchito maginito mikanda, mwambo laibulale ya kukula kwake angapezeke. ma buffers amatha kuwongolera mokhazikika komanso kuyesa magwiridwe antchito kuti akhazikike bwino komanso kuti azitha kuberekanso.
VAHTS Universal V8 RNA-seq Library Prep Kit ya Illumina itha kugwiritsidwa ntchito popanga RNA yokwanira ya poly(A) (ya RNA yokhala ndi umphumphu wabwino kuchokera ku eukaryotes monga zomera, nyama, ndi bowa) kapena rRNA yowononga laibulale ya RNA. Popeza kuti mRNA zomwe zili mu RNA yonse m'zitsanzo zosiyanasiyana zimasiyana kwambiri, ndikofunikira kuyika RNA yokwanira kuti mutsimikizire kuti mRNA yokwanira yomanga laibulale.
Ribo-off rRNA Depletion Kit (Munthu / Mouse / Rat) (Vazyme #N406): 0.01-1 g;VAHTS mRNA Tengani Mikanda (Vazyme #N401): 0.01-4 g;Ribo-off rRNA Depletion Kit (Bacteria) (Vazyme #N407): 0.01-5 g;Ribo-off rRNA Depletion Kit (chomera) (Vazyme #N409): 1-5 g;Ribo-off Globin & rRNA Depletion Kit (munthu / mbewa / makoswe) (Vazyme #N408): 0.01-1 g;mRNA yoyeretsedwa kapena ribosome yatha RNA: 0.5-100 ng.
Ndibwino kugwiritsa ntchito Agilent 2100 Bioanalyzer kuti muphunzire kukhulupirika kwa RNA.mRNA yonse yowonjezeredwa ndi VAHTS mRNA Capture Beads (Vazyme #N401) iyenera kukhala yapamwamba kwambiri ya RNA (RIN 7), monga RNA yowonongeka yogwiritsidwa ntchito pomanga laibulale ingayambitse 3 'Kukondera mu RNA-seq.Kwa zitsanzo za RNA zokhala ndi ma RIN <7, njira ya Ribo-off (Vazyme #N406/407/408/409) ndiyoyenera kuchotsa rRNA.
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