Kutevedzana kweRNA Diki-Illumina
Zvinhu zvirimo
● Kugadzirira raibhurari yekusarudza saizi.
● Ongororo yeBioinformatiki yakavakirwa pakufanotaura kwe miRNA uye zvinangwa.
Zvakanakira Basa
●Unyanzvi Hwakanyanya: Tagadzirisa sampuli dzinopfuura mazana matatu, dzichisanganisira mhando dzakasiyana dzesampuli. Tinounza hunyanzvi hwakawanda kupurojekiti yega yega.
●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.
●Kuongorora kwakazara kwebioinformatics:Inogonesa kuzivikanwa kwema miRNA anozivikanwa neatsva, kuzivikanwa kwezvinangwa zve miRNA, uye kutsanangurwa kwebasa rinoenderana uye kuwedzerwa nedhatabhesi dzakawanda (KEGG, GO).
●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 |
| Saizi yakasarudzwa | Illumina SE50 | 10M-20M inoverengwa | Q30≥85% |
Zvinodiwa zveMienzaniso:
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
Kutumirwa kwemuenzaniso kunokurudzirwa
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
● Kudzora kunaka kwedata
● Kurongwa kweRNA diki
● miRNA expression uye differential expression analysis
● gene rechinangwa chemiRNA uye gene rechinangwa chemiRNA rakatsanangurwa zvakasiyana-siyana
● Tsanangudzo yemajini anotarisirwa uye kuwedzera mashandiro emajini e miRNA anoratidzwa zvakasiyana
Kuzivikanwa kwe miRNA: chimiro uye kudzika
Kusiyanisa kwekutaura kwe miRNA - kuunganidzwa kwehiearchical
Tsanangudzo inoshanda yechinangwa che miRNA dzakaratidzwa zvakasiyana
Ongorora kufambira mberi kwekutsvaga kwakabatsirwa nemabasa eBMKGene ekutevera sRNA kuburikidza nemuunganidzwa wezvinyorwa zvakarongwa.
Chen, H. et al. (2023) 'Utachiona hwemavhairasi hunodzivirira saponin biosynthesis uye photosynthesis muPanax notoginseng',Physiology yezvirimwa neBiochemistry, 203, peji 108038. doi: 10.1016/J.PLAPHY.2023.108038.
Li, H. nevamwe (2023) 'Chirimwa chine mapuroteni FREE1 chinosanganisa zvinhu zve microprocessor kuti chidzivise miRNA biogenesis',Mishumo yeEMBO, 24(1). doi: 10.15252/EMBR.202255037/SUPPL_FILE/EMBR202255037-SUP-0004-SDATAFIG4.TIF.
Yu, J. et al. (2023) 'MicroRNA Ame-Bantam-3p Inodzora Kukura kweMapundu eLarval nekutarisa Multiple Epidermal Growth Factor-like Domains 8 Gene (megf8) muHoneybee, Apis mellifera',Chinyorwa Chepasi Pose cheSainzi dzeMolecular, 24(6), peji 5726. doi: 10.3390/IJMS24065726/S1.
Zhang, M. nevamwe (2018) 'Kuongororwa Kwakabatana kweMiRNA uye Majini Akabatana Nemhando Yenyama Kunoratidza kuti Gga-MiR-140-5p Inokanganisa Kuiswa Kwemafuta Mutsinga dzeHuku',Physiology yeMaseru neBiochemistry, 46(6), mapeji 2421–2433. doi: 10.1159/000489649.
