Sisebenzisa amakhukhi ukuthuthukisa ulwazi lwakho.Ngokuqhubeka upheqa lesi sizindalwazi uyavumelana nokusetshenziswa kwethu kwamakhukhi.Ulwazi olwengeziwe.
I-VAHTS Universal V8 RNA-seq Library Prep Kit ye-Illumina isetshenziselwa ukulungisa imitapo yolwazi ye-transcriptome yezinkundla ze-Illumina.Ikhithi iguquguquka futhi ifanele ukwakhiwa komtapo wezincwadi we-RNA we-RNA etholwe i-Poly(A)-based mRNA enrichment noma ukuncipha kwe-rRNA.
Ikhithi iqukethe izinhlobo ezimbili ze-cDNA 2nd Strand synthesis buffers. Ama-buffers angakhethwa ukwakha imitapo yolwazi yokuhlaziywa kwe-RNA-Seq transcriptome eboshiwe noma engagxilile. inqubo yokwakha umtapo wezincwadi futhi kunciphisa isikhathi sokusebenza.
Isistimu yokusabela ethuthukisiwe ithuthukisa ukusebenza kahle kokwakhiwa komtapo wolwazi, inikeza ukumbozwa okufanayo kwamanani ahlukene we-RNA yokufaka, futhi ihambisana nama-RNA okokufaka aphansi. Ngemva kokukhetha usayizi kusetshenziswa ubuhlalu obuzibuthe, kungatholwa umtapo wolwazi wangokwezifiso wosayizi othize.Ama-enzyme ekhithi kanye amabhafa angaphansi kokulawulwa kwekhwalithi okuqinile kanye nokuhlolwa kokusebenza kokuzinza okuphelele nokuphindaphindeka.
I-VAHTS Universal V8 RNA-seq Library Prep Kit ye-Illumina ingase isetshenziselwe ukwakha i-RNA ehlanganiswe ne-poly(A)-enothisiwe (ye-RNA yobuqotho obuhle evela kuma-eukaryote njengezitshalo, izilwane, nesikhunta) noma ukwakhiwa komtapo wezincwadi we-RNA wokuqeda i-rRNA. Njengoba okuqukethwe kwe-mRNA kwesamba se-RNA kumasampuli ahlukene kuhluka kakhulu, kuyadingeka ukufaka i-RNA ephelele eyanele ukuze kuqinisekiswe i-mRNA eyanele yokwakha umtapo wolwazi.
I-Ribo-off rRNA Depletion Kit (Umuntu/Igundane/Igundane) (Vazyme #N406): 0.01–1 g;I-VAHTS mRNA Thwebula Ubuhlalu (Vazyme #N401): 0.01–4 g;I-Ribo-off rRNA Depletion Kit (Amagciwane) (Vazyme #N407): 0.01–5 g;I-Ribo-off rRNA Depletion Kit (isitshalo) (Vazyme #N409): 1–5 g;I-Ribo-off Globin & rRNA Depletion Kit (umuntu/igundane/igundane) (Vazyme #N408): 0.01–1 g;i-mRNA ehlanzekile noma i-ribosome ephelile i-RNA: 0.5–100 ng.
Kungcono ukusebenzisa i-Agilent 2100 Bioanalyzer ukutadisha ubuqotho bengqikithi ye-RNA.mRNA ehambisana ne-VAHTS mRNA Capture Beads (Vazyme #N401) kufanele ibe i-RNA yekhwalithi ephezulu (RIN 7), njengoba isamba esibi esisetshenziselwe ukwakhiwa komtapo wolwazi singabangela u-3 ' ukuchema ku-RNA-seq.Kumasampuli e-RNA anamanani e-RIN <7, indlela ye-Ribo-off (Vazyme #N406/407/408/409) ifanele ukususwa kwe-rRNA.
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Isikhathi sokuthumela: May-06-2022