Ubude obupheleleyo be-mRNA Ukulandelelana-Nanopore
Iimbonakalo
● Ukuthatyathwa kwepoly-A mRNA elandelwa yi-cDNA synthesis kunye nokulungiswa kwethala leencwadi
● Ulandelelwano lwemibhalo ekhutshelweyo yobude obupheleleyo
● Uhlalutyo lwe-Bioinformatic olusekwe kulungelelwaniso lwe-reference genome
● Uhlalutyo lwe-Bioinformatiki alubandakanyi kuphela ukuchazwa kwejene kunye ne-isoform-level kodwa kunye nohlalutyo lwe-lncRNA, ukuxubana kwemfuza, i-poly-adenylation kunye nolwakhiwo lwemfuza.
Iinkonzo eziluncedo
●Ubungakanani bokubonakaliswa kwinqanaba le-isoform: ukwenza uhlalutyo oluneenkcukacha kunye noluchanekileyo lokuthetha, ukutyhila utshintsho olunokuthi lugqunywe xa kuhlalutywa yonke imbonakalo yemfuza
●IiMfuno eziNcitshisiweyo zeDatha:Xa kuthelekiswa ne-Next-Generation Sequencing (NGS), i-Nanopore sequencing ibonisa iimfuno ezisezantsi zedatha, ivumela amanqanaba alinganayo okulinganisa i-gene expression quantification saturation kunye nedatha encinci.
●Ukuchaneka okuphezulu kokuchazwa kobungakanani: zombini kwinqanaba lofuzo kunye ne-isoform
●Ukuchongwa kolwazi olongezelelweyo lwe-transcriptomic: enye i-polyadenylation, iijini ezidityanisiweyo kunye ne-lcnRNA kunye neentlobo zazo ekujoliswe kuzo
●UbuGcisa obuBanzi: Iqela lethu lizisa ubutyebi bamava kuyo yonke iprojekthi, sele ligqibe iiprojekthi ze-850 zeNanopore zobude obugcweleyo ze-transcriptome kwaye zisebenza ngaphezulu kwe-8,000 yeesampulu.
●Inkxaso yePost-Sales: Ukuzinikela kwethu kunabela ngaphaya kokugqitywa kweprojekthi kunye neenyanga ezi-3 emva kokuthengiswa kwenkonzo. Ngeli xesha, sinikezela ngokulandelelana kweprojekthi, uncedo lokusombulula ingxaki, kunye neeseshoni ze-Q&A ukujongana nayo nayiphi na imibuzo enxulumene neziphumo.
IiMfuno zeSampulu kunye nokuhanjiswa
| Ithala leencwadi | Isicwangciso sokulandelelana | Idatha iyacetyiswa | Ulawulo lwemeko |
| UPoly A utyebile | Illumina PE150 | 6/12 Gb | Amanqaku omgangatho ophakathi: Q10 |
IiMfuno zeSampulu:
IiNucleotides:
| Conc.(ng/μl) | Imali (μg) | Ubunyulu | Ingqibelelo |
| ≥ 100 | ≥ 1.0 | OD260/280=1.7-2.5 OD260/230=0.5-2.5 Unyino okanye akukho protein okanye ungcoliseko lweDNA olubonakaliswe kwijeli. | Kwizityalo: RIN≥7.0; Kwizilwanyana: RIN≥7.5; 5.0≥28S/18S≥1.0; umphakamo osisiseko okanye ongekhoyo |
● Izityalo:
Ingcambu, Isiqu okanye iPetal: 450 mg
Igqabi okanye Imbewu: 300 mg
Isiqhamo: 1.2 g
● Isilwanyana:
Intliziyo okanye Amathumbu: 300 mg
Viscera okanye Brain: 240 mg
Izihlunu: 450 mg
Amathambo, Iinwele okanye Ulusu: 1g
● IiArthropods:
Izinambuzane: 6g
I-crustacea: 300 mg
● Igazi elipheleleyo: ityhubhu e-1
● Iiseli: 106 iiseli
Unikezelo lweSampuli eCetyisiweyo
Isikhongozeli: 2 ml centrifuge ityhubhu (Ifoyile yeTin ayikhuthazwa)
Isampuli yokuleyibhela: Iqela+liphindaphinda umz. A1, A2, A3; B1, B2, B3.
Ukuthunyelwa ngenqanawa:
1. Umkhenkce owomileyo: Iisampulu kufuneka zipakishwe kwiingxowa kwaye zingcwatywe kumkhenkce owomileyo.
2. Iityhubhu ze-RNAstable: Iisampulu ze-RNA zingomiswa kwi-tube ye-RNA yokuzinzisa (umzekelo, i-RNAstable®) kwaye ithunyelwe kwiqondo lokushisa.
Ukuhamba komsebenzi weNkonzo
IiNucleotides:
Ukuhanjiswa kwesampuli
Ukwakhiwa kwethala leencwadi
Ukulandelelanisa
Uhlalutyo lwedatha
Iinkonzo emva kokuthengisa
Ukuhamba komsebenzi weNkonzo
Izicubu:
Uyilo lovavanyo
Ukuhanjiswa kwesampuli
Ukukhutshwa kwe-RNA
Ukwakhiwa kwethala leencwadi
Ukulandelelanisa
Uhlalutyo lwedatha
Iinkonzo emva kokuthengisa
● Ukusetyenzwa kwedatha ekrwada
● Ukuchongwa okubhaliweyo
● Enye indlela yokudibanisa
● Ubungakanani benkcazo kwinqanaba le-gene kunye nenqanaba le-isoform
● Uhlalutyo lwentetho eyahlukileyo
● Inkcazo yomsebenzi kunye nokutyebisa (iiDEGs kunye neeDETs)
Uhlalutyo olulolunye lwe-splicing
Uhlalutyo olulolunye lwePolyadenylation (APA)
Uqikelelo lwe-lncRNA
Inkcazo yemfuza yenoveli
Ukudityaniswa kwee-DETs
IiProtein-Protein Networks kwiiDEGs
Phonononga inkqubela phambili eququzelelwa yi-BMKGene's Nanopore yobude obugcweleyo beenkonzo zolandelelwano lwemRNA ngengqokelela egciniweyo yopapasho.
Gong, B. et al. (2023) 'I-Epigenetic kunye ne-transcriptional activation ye-secretory kinase FAM20C njenge-oncogene kwi-glioma', i-Journal ye-Genetics kunye ne-Genomics, i-50 (6), iphe. 422-433. doi: 10.1016/J.JGG.2023.01.008.
Yena, uZ. et al. (2023) 'Ukulandelelana kwe-transcriptome yobude obupheleleyo be-lymphocytes ukuphendula kwi-IFN-γ kubonisa i-Th1-skewed i-immune response in flounder (Paralichthys olivaceus)', I-Fish & Shellfish Immunology, i-134, iphe. 108636. doi: 10.1016/J.FSI.2023.108636.
UMa, Y. et al. (2023) 'Uhlalutyo oluthelekisayo lwePacBio kunye ne-ONT RNA yokulandelelana kweendlela zokuchongwa kwe-Nemopilema Nomurai venom', Genomics, 115 (6), p. 110709. doi: 10.1016/J.YGENO.2023.110709.
Yu, D. et al. (2023) 'Uhlalutyo lwe-Nano-seq lubonisa ukuthambekela okuhlukeneyo kokusebenza phakathi kwe-exosomes kunye ne-microvesicles ephuma kwi-hUMSC', i-Stem Cell Research and Therapy, i-14 (1), iphe. 1-13. doi: 10.1186/S13287-023-03491-5/ITABILE/6.
