Illumina horay loo sameeyay maktabadaha
Astaamaha
●Madal:Illumina NovaSeq 6000 iyo NovaSeq X Plus
●Hababka isku xigxiga:PE150 iyo PE250
●Xakamaynta tayada maktabadaha ka hor inta aan la isku dayin
●Isku xigxiga xogta QC iyo keenista:keenista warbixinta QC iyo xogta cayriin ee qaabka fastq ka dib kala saarista iyo shaandhaynta Q30 ayaa akhriya
Faa'iidooyinka Adeegga
●Baahsanaanta adeegyada isku xigxiga:macmiilku waxa uu dooran karaa isku xigxiga haadka, unugga socodka, ama cadadka xogta loo baahan yahay (isku xigxiga qaybta haadka).
●Khibrad ballaadhan oo ku saabsan goobta isku xigxiga ee Illumina:kumanaan mashruuc oo xidhan oo noocyo kala duwan leh.
●Soo gudbinta isku xigxiga warbixinta QC:oo leh cabbiro tayo leh, saxnaanta xogta iyo guud ahaan waxqabadka mashruuca isku xigxiga.
●Habka isku xigxiga ee qaangaadhka ah:oo leh waqti gaaban oo wareeg ah.
●Xakamaynta tayada adag: waxaan hirgelineynaa shuruudo adag oo QC ah si loo dammaanad qaado keenista natiijooyin tayo sare leh oo joogto ah.
Tusaalaha Platform
| Platform | Unugga socodka | Habka isku xigxiga | Unug | Wax soo saarka la qiyaasay |
| NovaSeq X | 10B (8 haad) | PE150 | Layn kali ah Layn Qaybeed | 375Gb / Layn |
| 25B (8 haad) | PE150 | Layn kali ah Layn Qaybeed | 1000 Gb/Lane | |
| NovaSeq 6000 | Unugga socodka SP (2 haad) | PE250 | Unugga socodka Layn kali ah Layn Qaybeed | 325-400 M akhrinta / Lane |
| Unugga socodka S4 (4 haad) | PE150 | Unugga socodka Layn kali ah Layn Qaybeed | ~ 800 Gb / Layn |
Shuruudaha Tusaalaha
| Qadarka Xogta (X) | Xooga saarida (qPCR/nM) | Mugga | |
| Taxanaha Qaybta Haadka
| X ≤ 10 Gb | ≥ 1 nM | ≥ 25 μl |
| 10 Gb <X ≤ 50 Gb | ≥ 2 nM | ≥ 25 μl | |
| 50 Gb <X ≤ 100 Gb | ≥ 3 nM | ≥ 25 μl | |
| X> 100 Gb | ≥ 4 nM | ≥ 25 μl | |
| Taxanaha Haadka | Layn kasta | ≥ 1.5 nM / barkadda maktabadda | ≥ 25 μl / barkadda maktabadda |
Marka lagu daro fiirsashada iyo wadarta qaddarka, qaab ku habboon ayaa sidoo kale loo baahan yahay.
Fiiro gaar ah: Isku xigxiga haadka ee maktabadaha kala duwanaanshiyaha hooseeya waxay u baahan tahay PhiX spike-in si loo xaqiijiyo wicitaanka sal adag.
Waxaan kugula talineynaa inaad soo gudbiso maktabado hore la isugu geeyay oo muunado ahaan ah. Haddii aad u baahan tahay BMKGENE si aad u sameyso isku dhafka maktabadda, fadlan tixraac
shuruudaha maktabadda ee taxanaha haadka qayb ahaan.
Cabbirka Maktabadda (Khariidadda ugu sarreysa)
Meesha ugu sarreysa waa in ay ahaataa 300-450 bp.
Maktabadu waa inay lahaadaan hal meel oo ugu sarreysa, oo aan lahayn wasakhda adabtarada iyo ma laha dimers-kii hore.
Fadlan nala soo xidhiidh haddii shaybaarkaagu aanu buuxin shuruudaha alaabta bilowga ah.
Socodka Shaqada Adeegga
Xakamaynta tayada maktabadda
Isku xigxiga
Xakamaynta tayada xogta
gaarsiinta mashruuca
Warbixinta maktabadda QC
Warbixin ku saabsan tayada maktabadda ayaa la bixiyaa ka hor inta aan la isku dayin, la qiimeynayo qaddarka maktabadda, iyo kala qaybsanaanta.
Isku xigxiga warbixinta QC
Shaxda 1. Tirakoobka xogta isku xigxiga.
| Tusaalaha Aqoonsiga | BMKID | Ceyriin wax akhriya | Xogta ceeriin (bp) | Akhris nadiif ah (%) | Q20 (%) | Q30 (%) | GC(%) |
| C_01 | BMK_01 | 22,870,120 | 6,861,036,000 | 96.48 | 99.14 | 94.85 | 36.67 |
| C_02 | BMK_02 | 14,717,867 | 4,415,360,100 | 96.00 | 98.95 | 93.89 | 37.08 |
Jaantuska 1. Qaybinta tayada oo la akhriyo muunad kasta
Jaantuska 2. Qaybinta nuxurka aasaasiga ah
Jaantuska 3. Qaybinta waxa la akhriyay ee xogta isku xigxiga
