Kutevedzana kweProkaryotic RNA
Zvinhu zvirimo
● Kugadziriswa kwesampuro yeRNA kwaisanganisira kudzikira kwe rRNA zvichiteverwa nekugadzirira raibhurari yeRNA nenzira.
● Ongororo yeBioinformatiki yakavakirwa pakuenderana ne genome reference
● Kuongorora kunosanganisira kuratidzwa kwemajini uye maDEG asiwo chimiro chezvinyorwa uye kuongorora kwesRNA
Zvakanakira Basa
●Kudzora Hunhu Kwakasimba: tinoshandisa nzvimbo dzekudzora zvinhu mumatanho ese, kubva pakugadzirira sampuro neraibhurari kusvika pakutevedzana uye bioinformatics. Kutarisa uku kwakanyatsonaka kunoita kuti pave nemhedzisiro yepamusoro nguva dzose.
●Data reKutevedzana kweStrand-specific: nekuda kwekuti gadziriro yeRNA raibhurari iri yekutungamira, zvichiita kuti zvikwanise kuzivikanwa kwezvinyorwa zvinopesana nepfungwa.
●Ongororo Yakazara Yakagadzirirwa Zvinyorwa zveProkaryotic: nzira ye bioinformatic haingosanganisiri kungoongorora mataurirwo emajini chete asiwo kuongorora chimiro chezvinyorwa, kusanganisira kuzivikanwa kwema operon, ma UTR uye zvinokurudzira. Inosanganisirawo kuongorora ma sRNA, kureva kutsanangura nekufanotaura chimiro chechipiri uye zvinangwa.
●Rutsigiro rweMushure meKutengesa: kuzvipira kwedu kunopfuura kupedzwa kwepurojekiti nemwedzi mitatu yebasa 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 | Maitiro ekuteverana | Data rinokurudzirwa | Kudzora Hunhu |
| raibhurari yenzira yakaderedzwa yerRNA | Illumina PE150 | 1-2 Gb | Q30≥85% |
Zvinodiwa zveMienzaniso:
| Conc.(ng/μl) | Huwandu (μg) | Kuchena | Kuvimbika |
| ≥ 50 | ≥ 1 | OD260/280=1.8-2.0 OD260/230=1.0-2.5 Kusvibiswa kwemapuroteni kana DNA kushoma kana kusina kuratidzwa pajeli. | RIN≥6.5 |
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
Kutumirwa kwemuenzaniso
Kuvakwa kweraibhurari
Kutevedzana kwezvikamu
Kuongorora data
Masevhisi ekupedzisira ekutengesa
Kufamba Kwebasa reKuongorora Bioinformatics
Inosanganisira Ongororo Inotevera:
● Kudzora kunaka kwedata
● Kuenderana ne genome reference
● Kuongorora Hunhu hweRaibhurari: Kupatsanurwa kweRNA kusingarongeki, saizi yekuisa uye kuzara kwezvikamu
● Tsanangudzo inoshanda yemajini ekunyora makodhi akafanotaurwa
● Kuongorora Matauriro: Kuwirirana uye Kuongorora Zvikamu Zvikuru (PCA)
● Kuratidzwa Kwemajini Akasiyana (DEGs)
● Tsanangudzo yekushanda uye kuwedzeredzwa kweDEGs
● Kuongorora kweRNA: kufanotaura, kududzira, chinangwa, uye kufanotaura kwechimiro chechipiri
● Kuongorora Chimiro cheTranscript: maoperon, nzvimbo dzekutanga nedzekupedzisira, Untranslated region (UTS), promoter, uye SNP/InDel analysis
Kutevedzana kwekuzara
Tsanangudzo inoshanda yemajini ekunyora makodhi
Kuwirirana pakati pemienzaniso
Kuongororwa kweMajini Akaratidza Kusiyana (DEGs)
Kuongorora kusimudzirwa kwebasa
tsananguro yesRNA
Ongorora kufambira mberi kuri kuitwa neBMKGene's Nanopore full-length mRNA sequencing services muchinyorwa chino chakakurumbira.
Guan, CP et al. (2018) 'Kuchinja kweTranscriptome yepasi rose kweBiofilm-Forming Staphylococcus epidermidis Kupindura kuTotal Alkaloids yeSophorea alopecuroides',Chinyorwa chePolish cheMicrobiology, 67(2), p. 223. doi: 10.21307/PJM-2018-024.
