Iilayibrari ezenziwe kwangaphambili ze-Illumina
Iimbonakalo
●Amaqonga:I-Illumina NovaSeq 6000 kunye neNovaSeq X Plus
●Iindlela zokulandelelanisa:I-PE150 kunye ne-PE250
●Ulawulo lomgangatho weelayibrari ngaphambi kokulandelelana
●Ukulandelelanisa idatha ye-QC kunye nokuhanjiswa kwayo:ukuhanjiswa kwengxelo ye-QC kunye nedatha eluhlaza kwifomathi ye-fastq emva kokususa izinto ezininzi kunye nokucoca ii-Q30 reads
Iingenelo zeNkonzo
●Ukuguquguquka kweenkonzo zokulandelelanisa:umthengi angakhetha ukulandelana ngomgca, iseli yokuhamba, okanye ngobungakanani bedatha efunekayo (ulandelelwano lwendlela engaphelelanga).
●Amava amaninzi kwiqonga lokulandelelana kwe-Illumina:kunye namawaka eeprojekthi ezivaliweyo ezineentlobo ngeentlobo.
●Ukuhanjiswa kwengxelo ye-QC yokulandelelana:ngemilinganiselo yomgangatho, ukuchaneka kwedatha kunye nokusebenza ngokubanzi kweprojekthi yokulandelelana.
●Inkqubo yokulandelelanisa abantu abadala:ngexesha elifutshane lokubuyela umva.
●Ulawulo Oluqinileyo Lomgangatho: Sisebenzisa iimfuno ze-QC ezingqongqo ukuqinisekisa ukunikezelwa kweziphumo ezisemgangathweni ophezulu rhoqo.
Iiplatfomu zeSampulu
| Iqonga | Iseli Yokuhamba | Imo yokulandelelanisa | Iyunithi | Imveliso eqikelelweyo |
| I-NovaSeq X | 10B (iindlela ezi-8) | PE150 | Indlela enye Indlela Engaphelelanga | 375Gb / Indlela |
| 25B (iindlela ezi-8) | PE150 | Indlela enye Indlela Engaphelelanga | 1000 Gb/Umgca | |
| I-NovaSeq 6000 | Iseli ye-SP Flow (iindlela ezi-2) | PE250 | Iseli Yokuhamba Indlela enye Indlela Engaphelelanga | Ufundo lwe-325-400 M / Indlela |
| Iseli ye-S4 Flow (iindlela ezi-4) | PE150 | Iseli Yokuhamba Indlela enye Indlela Engaphelelanga | ~800 Gb / Indlela |
Iimfuno zeSampuli
| Isixa seDatha (X) | Uxinaniso (qPCR/nM) | Umqulu | |
| Ulandelelwano Lwendlela Engaphelelanga
| 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 | |
| Ukulandelelana Kweendlela | NgokweNdlela nganye | ≥ 1.5 nM / Ichibi leThala leeNcwadi | ≥ 25 μl / Ichibi leThala leeNcwadi |
Ukongeza kuxinzelelo kunye nobungakanani obupheleleyo, kufuneka iphethini efanelekileyo yokuphakama.
Qaphela: Ukulandelelana kweendlela zeelayibrari ezinazo iintlobo ngeentlobo kufuna iPhiX inyuswe ukuze kuqinisekiswe ukuba isiseko sisebenza kakuhle.
Sicebisa ukuba singenise iilayibrari ezidityaniswe kwangaphambili njengeesampuli. Ukuba ufuna i-BMKGENE ukuba yenze i-library pooling, nceda ujonge ku
iimfuno zethala leencwadi zokulandelelana kweendlela ezingaphelelanga.
Ubungakanani beThala leeNcwadi (Imephu yeNcam)
Incopho ephambili kufuneka ibe phakathi kwe-300-450 bp.
Amathala eencwadi kufuneka abe ne-peak enye ephambili, kungabikho ungcoliseko lwe-adaptha kwaye kungabikho zi-primer dimers.
Nceda uqhagamshelane nathi ukuba iisampulu zakho azihlangabezani neemfuno zezinto zokuqala.
Ukuhamba komsebenzi wenkonzo
Ulawulo lomgangatho weThala leeNcwadi
Ukulandelelana
Ulawulo lomgangatho wedatha
Ukuhanjiswa kweprojekthi
Ingxelo yeQC yeThala leeNcwadi
Ingxelo ngomgangatho wethala leencwadi inikwa ngaphambi kokuba kulandelelaniswe, kuhlolwe ubungakanani bethala leencwadi, kwaye kuqhekeke.
Ingxelo ye-QC elandelelanayo
Itheyibhile 1. Izibalo malunga nedatha yokulandelelana.
| Isazisi seSampuli | I-BMKID | Ukufunda okungavuthwanga | Idatha eluhlaza (bp) | Ukufundwa okucocekileyo (%) | Q20(%) | Q30(%) | I-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 |
Umfanekiso 1. Ukusasazwa komgangatho ngokweempendulo ezifundwayo kwisampulu nganye
Umfanekiso 2. Ukusasazwa komxholo osisiseko
Umfanekiso 3. Ukusasazwa komxholo ofundiweyo kwidatha yokulandelelana
