I-V8 RNA-seq yokuLungiselela iKit yeThala leeNcwadi loLwakhiwo lweThala leencwadi le-RNA: Fumana iNtelekelelo, uBunde, iXabiso okanye Thenga

Sisebenzisa iikuki ukuphucula amava akho.Ngokuqhubeka ukhangela le sayithi uyavuma ukusebenzisa iikuki.Ingcaciso eyongezelelekileyo.
I-VAHTS Universal V8 RNA-seq ye-Library Prep Kit ye-Illumina isetyenziselwa ukulungiselela amathala eencwadi e-transcriptome kwiiplatifti ze-Illumina.Ikhithi iguquguqukayo kwaye ifanelekile kwi-RNA yethala leencwadi lokwakhiwa kwe-RNA efunyenwe yi-Poly (A)-based based mRNA enrichment okanye i-rRNA depletion.
Ikiti iqulethe iindidi ezimbini ze-cDNA 2nd Strand synthesis buffers.I-Buffers ingakhethwa ukwakha amathala eencwadi ukuze ihlalutye i-transcriptome ye-RNA-Seq ene-stranded okanye engaxilwanga. inkqubo yokwakha ithala leencwadi kunye nokunciphisa ixesha lokusebenza.
Inkqubo yokusabela ephuculweyo iphucula ukusebenza kakuhle kolwakhiwo lwethala leencwadi, ibonelela ngokujongwa ngokufanayo kwiimali ezahlukeneyo zegalelo le-RNA, kwaye iyahambelana nengeniso esezantsi ye-RNAs.Emva kokukhetha ubungakanani usebenzisa amaso amagnetic, ilayibrari yesiko lobungakanani obuthile inokufumaneka.I-enzymes zekhithi kunye izithinteli ziphantsi kolawulo olungqongqo lomgangatho kunye novavanyo olusebenzayo lozinzo olugqibeleleyo kunye nokuveliswa kwakhona.
I-VAHTS Universal V8 RNA-seq Prep Kit yeThala leeNcwadi ye-Illumina inokusetyenziselwa ukwakha nokuba yi-poly(A)-etyetyisiweyo iyonke i-RNA (ye-RNA yengqibelelo elungileyo evela kwi-eukaryotes efana nezityalo, izilwanyana, kunye nokungunda) okanye i-rRNA yokuncipha kokwakhiwa kwethala leencwadi le-RNA. Kuba umxholo we-mRNA we-RNA iyonke kwiisampulu ezahlukeneyo wahluka kakhulu, kuyimfuneko ukufaka i-RNA iyonke eyaneleyo ukuqinisekisa i-mRNA eyaneleyo kulwakhiwo lwethala leencwadi.
I-Ribo-off rRNA Kit Depletion Kit (Umntu / iMouse / Rat) (Vazyme #N406): 0.01-1 g;VAHTS mRNA Thatha amaso (Vazyme #N401): 0.01–4 g;I-Ribo-off rRNA Kit Depletion Kit (Bacteria) (Vazyme #N407): 0.01-5 g;I-Ribo-off rRNA Depletion Kit (isityalo) (Vazyme #N409): 1-5 g;I-Ribo-off Globin & rRNA Depletion Kit (umntu/impuku/impuku) (Vazyme #N408): 0.01–1 g;i-mRNA ecocekileyo okanye i-ribosome ephelelwe yi-RNA: 0.5-100 ng.
Kungcono ukusebenzisa i-Agilent 2100 Bioanalyzer ukufunda ukunyaniseka kwe-RNA.mRNA iyonke eyongezelelweyo nge-VAHTS mRNA Capture Beads (Vazyme #N401) kufuneka ibe yi-RNA ephezulu (RIN 7), njengoko i-RNA ethotyiweyo esetyenziselwa ukwakhiwa kwethala leencwadi ingabangela i-3. 'i-bias kwi-RNA-seq.Iisampuli ze-RNA ezinexabiso le-RIN <7, indlela ye-Ribo-off (Vazyme #N406/407/408/409) ifanelekile ukususwa kwe-rRNA.
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