Tixraaca aan ku salaysnayn mRNA Sequencing-Illumina
Astaamaha
● Ka madax banaan genome tixraac kasta,
● Xogta waxaa loo isticmaali karaa in lagu falanqeeyo qaabka iyo muujinta qoraallada
● Aqoonso goobaha goynta doorsooma
Faa'iidooyinka Adeegga
● gudbinta natiijada ku salaysan BMKCloud: Natiijooyinka waxaa loo dhiibaa sida faylka xogta iyo warbixinta isdhexgalka iyada oo loo sii marayo madal BMKCloud, taas oo u oggolaanaysa isticmaale-saaxiibtinimo akhrinta natiijooyinka falanqaynta adag iyo macdano la habeeyey oo ku salaysan falanqaynta caadiga ah ee bioinformatics.
● Adeegyada iibka kadib: 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
Tusaalaha Shuruudaha:
Nucleotides:
| Conc.(ng/μl) | Qadarka (μg) | daahirnimo | Daacadnimo |
| ≥ 20 | ≥ 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 |
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
Konteenar:
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.mRNA(denovo) Mabda'a Golaha
Saddexmidnimada, akhristuhu waa loo kala qaybiyaa qaybo yaryar, oo loo yaqaan K-mer.K-mer-yadan ayaa markaa loo isticmaalaa sidii iniin ahaan si loogu kordhiyo contigs ka dibna qayb ka mid ah iyadoo lagu saleynayo is-kordhinta is-kordhinta.Ugu dambeyntii, De Bruijn ayaa lagu dabaqay halkan si loo aqoonsado qoraallada qaybaha.
mRNA (De novo) Dulmarka Saddexmidnimada
2.mRNA (De novo) Qaybinta Heerka Muujinta Hida
RNA-Seq waxa ay awood u leedahay in ay gaadho qiyaas aad xasaasi u ah muujinta hiddesidaha.Caadiyan, tirada la ogaan karo ee muujinta qoraallada FPKM waxay u dhaxaysaa 10^-2 ilaa 10^6.
mRNA (De novo) Qaybinta cufnaanta FPKM muunad kasta
3.mRNA (De novo) GO Falanqaynta Kobcinta DEGs
Xogta GO (Gene Ontology) waa nidaam tafsiir bayooloji habaysan oo ka kooban erayo caadi ah oo hiddo-sidaha iyo shaqooyinka wax soo saarka hidda-wadaha.Waxay ka kooban tahay heerar kala duwan, halkaasoo heerka hoose uu yahay, inta badan hawlaha gaarka ah.
mRNA (De novo) GO kala soocida DEGs heerka labaad
Kiiska BMK
Falanqaynta Dhaqdhaqaaqa Dheef-shiid kiimikaadka Sukrose inta lagu jiro Bararka iyo Horumarinta Basasha (Allium cepa L.)
La daabacay: xuduudaha sayniska dhirta,2016
Istaraatiijiyada isku xigxiga
Illumina HiSeq2500
Tusaalaha ururinta
Beerta Jaallaha Isbaanishka ee Utah "Y1351" ayaa loo adeegsaday daraasaddan.Tirada shaybaarrada la qaaday waxay ahayd
Maalinta 15aad kadib bararka (DAS) ee nalka (dhexroorka 2-cm iyo miisaanka 3-4 g), 30aad DAS (dhexroorka 5-cm iyo miisaanka 100-110 g), iyo ~ 3 40-aad DAS (dhexroor 7-cm iyo 260-300 garaam).
Natiijooyinka muhiimka ah
1. jaantuska Venn, wadar ahaan 146 DEG ayaa laga helay dhammaan saddexda lamaane ee marxaladaha korriinka.
2.“Gaadiidka karbohaydraytka iyo dheef-shiid kiimikaadka” waxa metelayey 585 unugyo (ie, 7% COG-ga la sharraxay).
3.Unigenes si guul leh loogu sharraxay xogta GO ayaa loo kala saaray saddex qaybood oo muhiim ah saddexda marxaladood ee kala duwan ee horumarinta nalka.Inta badan waxa lagu matalay “habraaca bayoolojiga” qaybta ugu muhiimsan waxay ahaayeen “habsocodka dheef-shiid kiimikaadka”, oo ay ku xigto “habka gacanta”.Qaybta ugu muhiimsan ee "shaqada molecular" labada qaybood ee ugu badan ee lagu soo bandhigay waxay ahaayeen "binding" iyo "catalytic activity".
 Histogram of cluster of orthologous groups (COG) |  Heerarka hidda-socodka ontology (GO) ee kala soocida unigenes ee laga soo qaatay nalalka saddex marxaladood oo korriin ah |
 jaantuska Venn oo muujinaya hiddo-sideyaal si kala duwan loogu muujiyey laba marxaladood ee horumarinta guluubka basasha |
Tixraac
Zhang C, Zhang H, Zhan Z, iyo al.Falanqaynta Transcriptome ee Dheef-shiid kiimikaad Sukrose inta lagu jiro Bararka iyo Horumarka Bulb ee Basasha (Allium cepa L.)[J].Xuduudaha Sayniska Dhirta, 2016, 7:1425-.DOI: 10.3389/fpls.2016.01425
