Ulandelelwano lweTranscriptome epheleleyo – Illumina
Iimbonakalo
● Ukuveliswa kwethala leencwadi kabini ukuze kulandelelwe i-transcriptome epheleleyo: ithala leencwadi lokuphelelwa yi-rRNA kunye nokulungiswa kwethala leencwadi elincinci le-RNA.
● Uhlalutyo olupheleleyo lwe-bioinformatic lwe-mRNA, i-lncRNA, i-circRNA kunye ne-miRNA kwiingxelo ze-bioinformatic ezahlulweyo, kunye nohlalutyo oludibeneyo olune-RNA expression edibeneyo, kuquka nohlalutyo lweenethiwekhi ze-ceRNA.
Iingenelo zeNkonzo
●Ubuchule Obubanzi: Sisebenze iiprojekthi ze-transcriptome ezingaphezu kwama-2100, eziquka iintlobo ezahlukeneyo zeesampuli. Sizisa ubutyebi bobuchule kuyo yonke iprojekthi.
●Ulawulo Oluqinileyo Lomgangatho: Sisebenzisa amanqaku olawulo oluphambili kuzo zonke izigaba, ukusuka ekulungiseleleni iisampulu ukuya ekulungiseleleni ithala leencwadi, ukulandelelana kunye ne-bioinformatics. Ukubeka iliso kwethu ngononophelo kuqinisekisa ukunikezelwa kweziphumo ezisemgangathweni ophezulu rhoqo.
●Ingcaciso ebanzi: Sisebenzisa ii-database ezininzi ukuze sichaze ngokusebenzayo ii-Differentially Expressed Genes (DEGs) kwaye senze uhlalutyo lokufumisa oluhambelanayo. Le ndlela ibanzi inika ulwazi kwiinkqubo zeseli kunye neemolekyuli eziphantsi kwempendulo ye-transcriptome, ukuqinisekisa ukuba ufumana lonke ulwazi olunokwenzeka malunga nedatha yovavanyo lwakho.
●Uhlalutyo olunzulu lweeNethiwekhi zoLawuloUhlalutyo lwenethiwekhi ye-ceRNA lwenziwa ngokulandelelana okudibeneyo kwe-mRNA, i-lncRNA, i-circRNA, kunye ne-miRNA kunye nomsebenzi opheleleyo we-bioinformatic.
●Inkxaso Emva koThengiso: Siyaqonda ukubaluleka kokubakho, yiyo loo nto ukuzibophelela kwethu kudlulela ngaphaya kokugqitywa kweprojekthi kunye nexesha lenkonzo emva kokuthengiswa kweenyanga ezi-3. Ngeli xesha, sinikezela ngokulandela iprojekthi, uncedo lokusombulula iingxaki, kunye neeseshoni ze-Q&A ukuphendula nayiphi na imibuzo enxulumene neziphumo.
Iimfuno zeSampuli kunye nokuHanjiswa
| Ithala leeNcwadi | Icebo lokulandelelanisa izinto | Idatha iyacetyiswa | Ulawulo lwemeko |
| i-rRNA iphelile | I-Illumina PE150 | I-GB engama-16 | Q30≥85% |
| Ubungakanani obukhethiweyo | I-Illumina SE50 | Ukufundwa kwe-10-20M |
Iimfuno zeSampuli:
Iinukleotide:
| I-Conc.(ng/μl) | Ubungakanani (μg) | Ubunyulu | Ingqibelelo |
| ≥ 80 | ≥ 1.6 | OD260/280=1.7-2.5 OD260/230=0.5-2.5 Ungcoliseko lweproteni okanye lwe-DNA oluncinci okanye alubonakali kwijeli. | I-RIN≥6.0 5.0≥28S/18S≥1.0; ukuphakama okulinganiselweyo okanye okungekho siseko |
Ukuhanjiswa kweSampuli okuCetyisiweyo
Isikhongozeli: Ityhubhu ye-centrifuge ye-2 ml (Akukhuthazwa ukusebenzisa i-tin foil)
Iilebheli zesampulu: Iqela + ikopi umz. A1, A2, A3; B1, B2, B3.
Ukuthunyelwa:
1. Umkhenkce owomileyo: Iisampulu kufuneka zipakishwe kwiingxowa zize zingcwatywe kumkhenkce owomileyo.
2. Iityhubhu ze-RNAstable: Iisampulu ze-RNA zingomiswa kwityhubhu yokuzinzisa i-RNA (umz. i-RNAstable®) kwaye zithunyelwe kubushushu begumbi.
Ukuhamba komsebenzi wenkonzo
Uyilo lovavanyo
Ukuhanjiswa kwesampulu
Ukukhutshwa kwe-RNA
Ukwakhiwa kweThala leeNcwadi
Ukulandelelana
Uhlalutyo lwedatha
Iinkonzo zasemva kokuthengisa
I-Bioinformatics
Ngaphezu kwalo lonke uhlalutyo kwi-RNA nganye eyahlukileyo, iintlobo ezine ze-RNA zidityaniswe kuhlalutyo lwethu oludibeneyo:
- Uhlalutyo lokubonisana phakathi kwazo zonke iintlobo ezahlukeneyo zedatha ye-RNA
- Isishwankathelo sendlela yokubonakaliswa kwe-RNA
- Isishwankathelo sokwahluka kwembonakalo
- Inethiwekhi yeCeRNA
- Iijini eziphambili ezidityanisiweyo kwindlela
- Uhlalutyo lolondolozo
- Ubudlelwane obujolise kubo
Isishwankathelo sendlela yokubonakaliswa kwe-RNA
Izakhi zofuzo ezivezwe ngokwahlukileyo
Uhlalutyo lwe-ceRNA
Ii-miRNA ezivezwe ngokwahlukileyo kunye nee-RNA ezinxulumene nazo
Hlola inkqubela phambili yophando eququzelelwe ziinkonzo zokulandelelana kwe-BMKGene ngokusebenzisa ingqokelela yeempapasho ezikhethiweyo.
UDai, Y. et al. (2022) 'Iiprofayili zokubonakaliswa okubanzi kwee-mRNA, ii-lncRNA kunye nee-miRNA kwisifo sikaKashin-Beck ezichongiweyo ngokulandelelana kwe-RNA',Ii-Molecular Omics, 18(2), iphepha 154-166. doi: 10.1039/D1MO00370D.
ULiu, N. nan et al. (2022) 'Uhlalutyo olupheleleyo lwe-transcriptomes yokumelana nokubanda kwe-Apis cerana kwiNtaba yaseChangbai ngexesha lobusika.',Ijini, 830, iphe. 146503–146503. doi: 10.1016/J.GENE.2022.146503.
UWang, XJ et al. (2022) 'Ukubekwa phambili okusekwe kwi-Multi-Omics Integration-Based Prioritisation of Competing Endogenous RNA Regulation Networks in Small Cell Lung Cancer: Iimpawu zeMolecular kunye naBafaki beZiyobisi',Imida kwi-Oncology, 12, iphe. 904865. doi: 10.3389/FONC.2022.904865/BIBTEX.
UXu, P. et al. (2022) 'Uhlalutyo oludibeneyo lweeprofayili zokubonakaliswa kwe-lncRNA/circRNA-miRNA-mRNA lutyhila ukuqonda okutsha malunga neendlela ezinokubakho zokuphendula kwi-root-knot nematodes kwi-peanut',I-BMC Genomics, 23(1), iphe. 1–12. doi: 10.1186/S12864-022-08470-3/FIGURES/7.
UYan, Z. et al. (2022) 'Ulandelelwano lwe-RNA olupheleleyo lugxininisa iindlela ze-molecular ezinxulumene nokugcinwa komgangatho emva kokuvunwa kwi-broccoli nge-LED irradiation ebomvu',Ibhayoloji kunye neTekhnoloji emva kokuvuna, 188, iphe. 111878. doi: 10.1016/J.POSTHARIVBIO.2022.111878.
