Ukulandelelana kweCircRNA-Illumina
Iimbonakalo
● Ukuncipha kwe-rRNA kulandelwa kukulungiswa kwethala leencwadi elikhokelayo, okuvumela idatha yokulandelelana kwemigca ethile.
● Umsebenzi we-Bioinformatic uvumela ukuqikelela kwe-circRNA kunye nokulinganisa ukubonakaliswa.
Iingenelo zeNkonzo
●Ubuchule Obubanzi: Sicubungule iisampulu ezingaphezu kwama-20,000, sithatha iintlobo ezahlukeneyo zeesampulu, 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.
●IiLayibrari zeRNA eziBanzi ngakumbi:Sisebenzisa i-rRNA depletion endaweni ye-linear RNA depletion kulungiselelo lwethu lwangaphambi kwethala leencwadi, siqinisekisa ukuba idatha yokulandelelana ayibandakanyi kuphela i-circRNA kodwa kunye ne-mRNA kunye ne-lncRNA, okuvumela uhlalutyo oludibeneyo lwezi seti zedatha.
●Uhlalutyo oluKhethekileyo lweeNethiwekhi ze-RNA (ceRNA) eziKhuphisanayo: Ukubonelela ngolwazi olunzulu malunga neendlela zokulawula iiseli.
● Inkxaso Emva kokuthengisa: 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 | Iqonga | Idatha ecetyiswayo | Idatha ye-QC |
| ilayibrari yesikhokelo ephelileyo ye-rRNA | I-Illumina PE150 | 16-20 Gb | Q30≥85% |
Iimfuno zeSampuli:
Iinukleotide:
| I-Conc.(ng/μl) | Ubungakanani (μg) | Ubunyulu | Ingqibelelo |
| ≥ 80 | ≥ 0.8 | 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 |
● Izityalo:
Ingcambu, Isiqu okanye iPetal: 450 mg
Igqabi okanye Imbewu: 300 mg
Isiqhamo: 1.2 g
● Isilwanyana:
Intliziyo okanye Amathumbu: 450 mg
I-Viscera okanye ubuchopho: 240 mg
Izihlunu: 600 mg
Amathambo, Iinwele okanye Ulusu: 1.5g
● Ii-arthropod:
Izinambuzane: 9g
I-Crustacea: 450 mg
● Igazi elipheleleyo:Iityhubhu ezi-2
● Iiseli: 106 iiseli
● I-Serum kunye nePlasma: 6 mL
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
- Idatha ye-QC
- Ulungelelwaniso lwejini
- Ukuchongwa kwe-CirRNA
- Ukubonakaliswa kwe-CirRNA
- Uhlalutyo lokubonakaliswa kokwahluka
Uqikelelo lwe-circRNA: usasazo lwe-chromosome
Ii-circRNA ezichazwe ngokwahlukileyo – iploti yevolcano
Ii-circRNA ezichazwe ngokwahlukileyo - ukuhlanganiswa kwe-hierarchical
Ukutyebisa okusebenzayo kwezakhi zofuzo ze-circRNA
Hlola inkqubela phambili yophando eququzelelwe ziinkonzo zokulandelelana kwe-BMKGene' circRNA ngokusebenzisa ingqokelela yeempapasho ezikhethiweyo.
UWang, X. et al. (2021) 'I-CPSF4 ilawula ukwakheka kwe-circRNA kunye nokuthuliswa kwezakhi zofuzo ezidityaniswe yi-microRNA kwi-hepatocellular carcinoma',I-Oncogene2021 40:25, 40(25), iphe. 4338–4351. doi: 10.1038/s41388-021-01867-6.
UXia, K. et al. (2023) 'I-X oo-responsive transcriptome ityhila indima ye-RNA133 ejikelezayo ekuchaseni izifo ngokulawula ukubonakaliswa kwe-OsARAB kwirayisi',Uphando lweFitopathology, 5(1), iphe. 1–14. doi: 10.1186/S42483-023-00188-8/FIGURES/6.
Y, H. et al. (2023) 'I-CPSF3 ilungisa ibhalansi yeengxelo ezijikelezayo nezithe ngqo kwi-hepatocellular carcinoma'. doi: 10.21203/RS.3.RS-2418311/V1.
UZhang, Y. et al. (2023) 'Uvavanyo olupheleleyo lwee-circRNAs kwi-cirrhotic cardiomyopathy ngaphambi nasemva kokufakelwa kwesibindi',I-Immunopharmacology yaMazwe ngaMazwe, 114, iphe. 109495. doi: 10.1016/J.INTIMP.2022.109495.
