Kutevedzana Kwenguva Refu Kusingakodhese Makodhi-Illumina
Zvakanakira Basa
●Kuongororwa kwakabatana kwemRNA nelncRNA: nekubatanidza kuverengwa kwezvinyorwa zvemRNA nekudzidza kwelncRNA nezvinangwa zvavo, zvinokwanisika kuwana ruzivo rwakadzama rwenzira yekudzora iri pasi pemhinduro yemasero.
●Unyanzvi Hwakanyanya: Tagadzirisa sampuro dzinopfuura 230,000, dzichisanganisira sampuli dzakasiyana-siyana uye zvinangwa zvemapurojekiti. Tinounza hunyanzvi hwakawanda kupurojekiti yega yega.
●Kuongororwa kwakabatana kwemRNA nelncRNA: Tinosanganisa huwandu hwezvinyorwa zvemRNA nekudzidza kwelncRNA nezvinangwa zvavo, zvichiita kuti tikwanise kuwana ruzivo rwakadzama rwemaitiro ekutonga ari pasi pemhinduro yemasero.
●Kudzora Hunhu Kwakasimba: Tinoshandisa nzvimbo dzekudzora zvinhu mumatanho ese, kubva pakugadzirira sampuro kusvika pakugadzirira raibhurari, kutevedzana kwezvinhu uye bioinformatics. Kutarisa kwedu kwakanyatsonaka kunoita kuti pave nemhedzisiro yepamusoro nguva dzose.
●Tsanangudzo Yakazara: Tinoshandisa madhatabhesi akawanda kuti titsanangure mashandiro eDifferentially Expressed Genes (DEGs) uye kuita ongororo dzinoenderana nekuwedzera ruzivo. Iyi nzira yakazara inopa ruzivo mumabasa emasero nemamorekuru ari pasi pemhinduro yetranscriptome, zvichiita kuti uwane ruzivo rwese rwunogoneka nezve data rekuyedza kwako.
●Rutsigiro rweMushure meKutengesa: Tinonzwisisa kukosha kwekuvapo, ndosaka kuzvipira kwedu kusingapfuuri kupedzwa kweprojekiti nenguva yebasa remwedzi mitatu mushure mekutengesa. Munguva ino, tinopa rubatsiro rwekutevera purojekiti, rubatsiro rwekugadzirisa matambudziko, uye mibvunzo nemhinduro kuti tipindure chero mibvunzo ine chekuita nemhedzisiro.
Zvinodiwa zveMienzaniso uye Kutumirwa
| Raibhurari | Puratifomu | Ruzivo runokurudzirwa | QC yedata |
| raibhurari yenzira yakaderedzwa yerRNA | Illumina PE150 | 10-16 Gb | Q30≥85% |
Manyukireotidhi:
| Conc.(ng/μl) | Huwandu (μg) | Kuchena | Kuvimbika |
| ≥ 80 | ≥ 0.8 | OD260/280=1.7-2.5 OD260/230=0.5-2.5 Kusvibiswa kwemapuroteni kana DNA kushoma kana kusina kuratidzwa pajeli. | RIN≥6.0; 5.0≥28S/18S≥1.0; kukwirira kwepakutanga kwakaganhurirwa kana kusina |
● Zvirimwa:
Mudzi, Hunde kana Petal: 450 mg
Shizha kana Mbeu: 300 mg
Muchero: 1.2 g
● Mhuka:
Moyo kana Utumbo: 450 mg
Viscera kana Uropi: 240 mg
Tsandanyama: 600 mg
Mapfupa, Bvudzi kana Ganda: 1.5g
● Mapodhi emhuka:
Zvipembenene: 9g
Chibage cheCrustacea: 450 mg
● Ropa rose:Machubhu maviri
● Masero: 106 masero
● Serum nePlasma: 6 mL
Muenzaniso Unokurudzirwa Kutumirwa
Chirongo: 2 ml chubhu ye centrifuge (Haikurudzirwi kuisa tin foil)
Kunyora muenzaniso: Boka + dzokorora semuenzaniso A1, A2, A3; B1, B2, B3.
Kutumirwa:
1. Aizi yakaoma: Mienzaniso inofanira kurongedzwa mumabhegi uye kuvigwa muaizi yakaoma.
2. Machubhu eRNAstable: Samples dzeRNA dzinogona kuomeswa muchubhu yeRNA stabilization (semuenzaniso RNAstable®) uye dzinotumirwa mutembiricha yemumba.
Kufamba Kwebasa Rebasa
Dhizaini yekuedza
Kutumirwa kwemuenzaniso
Kubviswa kweRNA
Kuvakwa kweraibhurari
Kutevedzana kwezvikamu
Kuongorora data
Masevhisi ekupedzisira ekutengesa
Bioinformatics
- Ruzivo rusina kubikwa
- QC yedata
- Kurongeka kwemajini
- Maumbirwo emajini (Kumwe kusanganisa majini, kugadzirisa maumbirwo emajini uye kufanotaura kwemajini matsva)
- Kuyera kwekutaura kwemajini
- Kuongorora kusiyana kwekutaura
- Kutsanangurwa kweDEG uye kusimudzirwa + majini echinangwa cheLNCRNA anoratidzwa zvakasiyana
- Kuzivikanwa kwechinyorwa
- Kuzivikanwa kweLncRNA (kuchengetedzwa kweLncRNA uye kuzivikanwa kweLncRNA)
- Kufanotaura kwemajini echinangwa chelncRNA
- Kuyera kwekutaura kwelncRNA
- Kuongororwa pamwe chete nedata re mRNA
Kuongorora kweDifferential Gene Expression (DEGs)
Kuyera kwekutaura kwelncRNA - kuunganidzwa
Kuwedzerwa kwemajini echinangwa chelncRNA
Kuongorora nzvimbo yemRNA pamwe chete uye lncRNA - Circos plot (denderedzwa repakati i mRNA uye inner cicrlce i lncRNA)
Ongorora kufambira mberi kuri kuitwa neBMKGene's lncRNA equation services kuburikidza nemuunganidzwa wezvinyorwa zvakarongwa.
Ji, H. et al. (2020) 'Kuzivikanwa, kufanotaura mashandiro, uye kusimbiswa kwakakosha kweLNCRNA kweLNCRNA dzine chekuita nekushushikana muchiropa chemakonzo',Mishumo yeSainzi2020 10:1, 10(1), mapeji 1–14. doi: 10.1038/s41598-020-57451-7.
Jia, Z. nevamwe (2021) 'Kuongororwa kweTranscriptomic Integrative Kunoratidza Maitiro Ekudzivirira Muviri weCyHV-3-Resistant Common Carp Strain',Miganhu muImmunology, 12, p. 687151. doi: 10.3389/FIMMU.2021.687151/BIBTEX.
Wang, XJ nevamwe (2022) 'Kubatanidzwa kweMulti-Omics-Based Prioritisation yeEndogenous RNA Regulation Networks muDiki Cell Lung Cancer: Hunhu hweMolecular uye Vanogona Kutora Mishonga',Miganhu muOncology, 12, p. 904865. doi: 10.3389/FONC.2022.904865/BIBTEX.
Xiao, L. et al. (2020) 'Kupatsanurwa kwemajini kwenetiweki yekubatana kwemajini iri pasi pe photosynthesis muPopulus',Chinyorwa cheBiotechnology yezvirimwa, 18(4), mapeji 1015–1026. doi: 10.1111/PBI.13270.
Zheng, H. nevamwe (2022) 'Nhare Yenyika Yose Yekudzora Kubuda Kwemajini Asina Kurongeka uye Kuratidza Kusagadzikana Kwemetabolism muMaseru Ekudzivirira Muviri muChirwere CheGraves' uye Hashimoto's Thyroiditis',Miganhu muImmunology, 13, p. 879824. doi: 10.3389/FIMMU.2022.879824/BIBTEX.
