Xigasho dheer oo aan codayn-Ilumina
Faa'iidooyinka Adeegga
● Faa'iidooyinka Adeegga
● Unugyo gaar ah iyo unugyo gaar ah
Marxaladda gaarka ah waxay muujisaa oo soo bandhigtaa isbeddelka odhaahda firfircoon
● Hababka saxda ah ee wakhtiga iyo hadalka goobta
● Falanqaynta wadajirka ah ee xogta mRNA.
● Gaadhista natiijada ku salaysan BMKCloud: Macdanta habaysan ee laga heli karo goobta.
● Adeegyada iibka kadib waxay shaqaynayaan 3 bilood marka mashruuca la dhammeeyo
Tusaalaha Shuruudaha iyo Bixinta
| Maktabadda | Platform | Xogta lagu taliyay | Xogta QC |
| dhimista rRNA | Illumina PE150 | 10 Gb | Q30≥85% |
| Conc.(ng/μl) | Qadarka (μg) | daahirnimo | Daacadnimo |
| ≥ 100 | ≥ 0.5 | OD260/280=1.7-2.5 OD260/230=0.5-2.5 Xaddidan ama aan lahayn borotiin ama wasakh DNA oo lagu muujiyey jel. | Dhirta: RIN≥6.5; Xoolaha: RIN≥7.0; 5.0≥28S/18S≥1.0; xaddidan ama aan lahayn sare u qaadis aasaasi ah |
Nucleotides:
Nudaha: Miisaanka (qallalan): ≥1 g
* Unugyada ka yar 5 mg, waxaan kugula talineynaa inaad soo dirto muunada nudaha barafaysan (ku jira nitrogen dareere ah).
Ka joojinta unugga: Tirada unugyada = 3×107
* Waxaan kugula talineynaa in la raro unugyada la qaboojiyey.Haddii unuggaasi uu ka yar yahay 5×105, Tooshka barafaysan ee dareeraha nitrogen ayaa lagula talinayaa.
Muunadaha dhiigga:
PA × geneBloodRNATube;
6mLTRIzol iyo 2ml dhiig (TRIzol:Blood=3:1)
Tusaalaha keenista lagu taliyay
Weel: 2 ml oo tuubo sentrifuge ah (Faylada qasacadaysan laguma talinayo)
Tusaalaha calaamadaynta: Kooxda+ku celcelinta tusaale A1, A2, A3;B1, B2, B3......
Raritaanka:
1.Baraf qallalan: Tijaabooyinku waxay u baahan yihiin in lagu xidho bacaha oo lagu aaso baraf qallalan.
Tubooyinka 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
1.LncRNA kala saarista
LncRNA oo ay saadaaliyeen afarta softwares ee kor ku xusan ayaa loo kala saaray 4 qaybood: lincRNA, anti-sense-LncRNA, intronic-LncRNA;dareen-LncRNA.Kala soocida LncRNA waxaa lagu muujiyay Histogramka hoose.
Kala soocidda LncRNA
2.Cis-bartilmaameedka hiddo-wadaha ee falanqaynta kobcinta DE-lncRNA
ClusterProfiler waxaa lagu shaqaaleysiiyay falanqaynta kobcinta GO ee hiddo-wadaha cis-ku wajahan ee lncRNA (DE-lncRNA) si ka duwan loo muujiyay, marka la eego hababka noolaha, hawlaha molecular iyo qaybaha gacanta.Falanqaynta kobcinta GO waa hab lagu aqoonsado kelmadaha GO ee ay DEG-dugeyso oo aad loo tayeeyey marka la barbar dhigo genome oo dhan.Erayada la hodmay ayaa lagu soo bandhigay histogram, shaxda xumbo, iwm. sida hoos ku cad.
Hidde-sideyaasha Cis-ku-talagalka ah ee falanqaynta kobcinta DE-lncRNA - shaxda xumbo
3. Marka la barbardhigo dhererka, nambarka exon, ORF iyo qadarka muujinta mRNA iyo lncRNA, waxaan fahmi karnaa kala duwanaanshaha qaab dhismeedka, isku xigxiga iyo wixii la mid ah dhexdooda, sidoo kale waxaan hubin karnaa in lncRNA novel ee annagu saadaaliyay uu waafaqsan yahay sifooyinka guud.
Kiiska BMK
Tilmaamaha muujinta lncRNA ee la dhimay ee adenocarcinomas sambabada jiirka oo leh isbeddelka KRAS-G12D iyo garaacista P53
La daabacay:Wargeyska Daawaynta Unugyada iyo Unugyada2019
Istaraatiijiyada isku xigxiga
Illumina
Tusaalaha ururinta
Unugyada NONMMUT015812-knockdown KP (shRNA-2) iyo unugyada xakamaynta taban (sh-Scr) ayaa la helay maalinta 6 ee caabuqa fayraska gaarka ah.
Natiijooyinka muhiimka ah
Daraasadani waxay baari doontaa lncRNA-yada si xun loo muujiyey ee adenocarcinoma sambabada jiirka oo leh garaaca P53 iyo isbeddelka KrasG12D.
1.6424 lncRNAs si kala duwan ayaa loo muujiyay (≥ 2-laab beddel, P <0.05).
2.Dhammaan 210 lncRNAs (FC≥8), 11 lncRNAs 'muujinta waxaa nidaamiyay P53, 33 lncRNAs by KRAS iyo 13 lncRNAs by hypoxia ee unugyada KP aasaasiga ah, siday u kala horreeyaan.
3.NONMMUT015812, kaas oo si cajiib ah kor loogu habeeyay adenocarcinoma sambabada jiirka oo si xun u nidaamiyay P53 dib-u-muujinta, ayaa la ogaaday si loo falanqeeyo shaqadiisa gacanta.
4.Knockdown ee NONMMUT015812 by shRNAs waxay hoos u dhigtay awoodaha iyo guuritaanka unugyada KP.NONMMUT015812 waxay ahayd konjine.
 Falanqaynta dariiqa KEGG ee hiddo-wadaha si ka duwan loo muujiyey ee ku jira unugyada NONMMUT015812-knockdown KP |  Falanqaynta Gene Ontology ee hiddo-wadaha si ka duwan loo muujiyey ee ku jira unugyada NONMMUT015812-knockdown KP |
Tixraac
Tilmaamaha muujinta lncRNA ee la dhimay ee adenocarcinomas sambabada jiirka oo leh isbeddelka KRAS-G12D iyo garaacista P53 [J].Journal of Cellular iyo Molecular Medicine, 2019, 23(10).DOI: 10.1111/jcmm.14584
