Eukaryotic mRNA isku xigxiga-Illumina
Faa'iidooyinka
ØKhibrad sare leh: In ka badan 200,000 oo muunado ayaa lagu farsameeyay BMK oo daboolaya noocyo kala duwan oo muunado ah, oo ay ku jiraan dhaqanka unugyada, unugyada, dareeraha jidhka, iwm. iyo in ka badan 7,000 mRNA-Seq mashruucyo ayaa la xidhay oo daboolaya aag cilmi-baaris oo kala duwan.
ØNidaamka xakamaynta tayada adag: Qodobbada xakamaynta tayada asaasiga ah iyada oo loo marayo dhammaan tallaabooyinka ay ku jiraan diyaarinta muunada, diyaarinta maktabadda, isku xigxiga iyo bioinformatics ayaa si dhow loola socdaa si loo bixiyo natiijooyin tayo sare leh.
ØXog ururin badan oo diyaar u ah sharraxaadda shaqada iyo daraasadaha kobcinta si loo fuliyo ujeedooyinka cilmi-baarista ee kala duwan.
ØAdeegyada iibka ka dib: Adeegyada iibka kadib waxay shaqaynayaan 3 bilood marka mashruuca la dhammeeyo, oo ay ku jiraan dabagalka mashaariicda, dhib-toogashada, natiijooyinka Q&A, iwm.
Tusaalaha Shuruudaha iyo Bixinta
| Maktabadda | Istaraatiijiyada isku xigxiga | Xogta lagu taliyay | Xakamaynta tayada |
| Poly A waa la hodmay | Illumina PE150 | 6 Gb | Q30≥85% |
Tusaalaha Shuruudaha:
Nucleotides:
| daahirnimo | Daacadnimo | Qadarka |
| OD260/280≥1.7-2.5 OD260/230≥0.5-2.5Xadidan ama aan lahayn wasakhowga borotiinka ama DNA-ga ee lagu muujiyey jel. | Dhirta: RIN≥6.5; Xayawaanka: RIN≥7;28S/18S≥1.0; xadidan ama aan sal lahayn | Conc.≥30 ng/μl;Volum ≥ 10 μl;Wadarta ≥ 1.5 μg |
Nudaha: Miisaanka (qallalan):≥1 g
*Wixii ka yar 5 mg, waxaan kugula talineynaa inaad soo dirto muunada nudaha barafaysan (ku jira nitrogen dareere ah).
Ka joojinta unugga:Tirada unugyada = 3×106- 1×107
* Waxaan kugula talineynaa in la soo dejiyo lysate unug barafaysan.Haddii unugyadu ay ka yar yihiin 5 × 105, fiilo barafaysan oo dareere ah ayaa lagula talinayaa, taas oo u fiican soo saarista yar yar.
Muunadaha dhiigga:Mugga ≥1 ml
Microorganism:Cuf ≥ 1 g
Tusaalaha keenista lagu taliyay
Weel: 2 ml tuubo sentrifuge (Faylada qasacadaysan laguma talinayo)
Tusaalaha calaamadaynta: Kooxda+ku celcelinta tusaale A1, A2, A3;B1, B2, B3......
Raritaanka:
- Barafka qallalan: Muunada waxay u baahan yihiin in lagu soo raro bac oo lagu aaso baraf qallalan.
- Tuubooyinka RNAstable: Muunada RNA waxaa lagu qallajin karaa tuubada xasilinta RNA (tusaale RNAstable®) waxaana lagu rari karaa heerkulka qolka.
Socodka Shaqada Adeegga
Naqshadaynta tijaabada
Tusaalaha keenista
soo saarista RNA
Dhismaha maktabadda
Isku xigxiga
Falanqaynta xogta
Adeegyada iibka kadib
Bioinformatics
Eukaryotic socodka shaqada falanqaynta taxanaha mRNA
Bioinformatics
ØXakamaynta tayada xogta ceeriin
ØTixraaca genome alignment
ØFalanqaynta qaab-dhismeedka qoraalka
ØQiyaasta muujinta
ØFalanqaynta odhaah kala duwan
ØSharaxaada shaqada iyo kobcinta
1.mRNA Xogta Saturation curve
2.Falanqaynta tibaaxaha kala duwan -Dhulka Volcano
3.Faahfaahinta KEGG ee DEGs
4.Kala soocida GO ee DEGs
