Ukulandelelana kwe-RNA encinci-Illumina
Iimbonakalo
● Ukulungiselela ithala leencwadi lokukhetha ubungakanani.
● Uhlalutyo lwe-Bioinformatic lujolise ekuqikeleleni i-miRNA kunye neethagethi.
Iingenelo zeNkonzo
●Ubuchule Obubanzi: Sicubungule iisampulu ezingaphezu kwama-300, eziquka 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.
●Uhlalutyo olupheleleyo lwe-bioinformatics:Ivumela ukuchongwa kwee-miRNA ezaziwayo nezintsha, ukuchongwa kweethagethi ze-miRNA, kunye nokuchazwa okusebenzayo kunye nokuphuculwa ngeedathabheyisi ezininzi (KEGG, GO).
●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 | Iqonga | Idatha ecetyiswayo | Idatha ye-QC |
| Ubungakanani obukhethiweyo | I-Illumina SE50 | Ukufundwa kwe-10M-20M | 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
Ukukhupha i-RNA
Ukwakhiwa kweThala leeNcwadi
Ukulandelelana
Uhlalutyo lwedatha
Iinkonzo zasemva kokuthengisa
I-Bioinformatics
● Ulawulo lomgangatho wedatha eluhlaza
● Ukwahlulwahlulwa kwe-RNA encinci
● uhlalutyo lokubonakaliswa kwe-miRNA kunye nolwahlulo lokubonakaliswa kokwahluka
● i-gene ekujoliswe kuyo ye-miRNA kunye ne-gene ekujoliswe kuyo ye-miRNA echazwe ngokwahlukileyo
● Inkcazelo yejini ekujoliswe kuyo kunye nokuphuculiswa komsebenzi wejini ye-miRNA echazwe ngokwahlukileyo
Ukuchongwa kwe-miRNA: isakhiwo kunye nobunzulu
Ukubonakaliswa okwahlukileyo kwe-miRNA - ukuhlanganiswa kwe-hiearchical
Isichazi-magama esisebenzayo senjongo yee-miRNA ezivezwe ngokwahlukileyo
Hlola inkqubela phambili yophando eququzelelwe ziinkonzo zokulandelelana kwe-sRNA ze-BMKGene ngokusebenzisa ingqokelela yeempapasho ezikhethiweyo.
UChen, H. et al. (2023) 'Usulelo lwentsholongwane luthintela i-saponin biosynthesis kunye ne-photosynthesis kwiPanax notoginseng',I-Physiology yeZityalo kunye neBiochemistry, 203, iphe. 108038. doi: 10.1016/J.PLAPHY.2023.108038.
ULi, H. et al. (2023) 'Isityalo esine-FYVE domain-containing protein FREE1 sinxulumana nezinto ze-microprocessor ukuze kuthintelwe i-miRNA biogenesis',Iingxelo ze-EMBO, 24(1). doi: 10.15252/EMBR.202255037/SUPPL_FILE/EMBR202255037-SUP-0004-SDATAFIG4.TIF.
Yu, J. et al. (2023) 'I-MicroRNA Ame-Bantam-3p ilawula uphuhliso lwe-Larval Pupal ngokujolisa kwi-Multiple Epidermal Growth Factor-like Domains 8 Gene (megf8) kwi-Honeybee, Apis mellifera',Ijenali yeHlabathi yeSayensi yeMolekyuli, 24(6), iphe. 5726. doi: 10.3390/IJMS24065726/S1.
UZhang, M. et al. (2018) 'Uhlalutyo oluDibeneyo lweMiRNA kunye neeGene eziDibene noMgangatho weNyama lutyhila ukuba iGga-MiR-140-5p ichaphazela ukubola kwamafutha ngaphakathi kwezihlunu kwiinkukhu',I-Cellular Physiology kunye neBiochemistry, 46(6), iphepha 2421–2433. doi: 10.1159/000489649.
