Iimveliso

Illumina kunye ne-BGI

Itekhnoloji yolandelelwano ye-Illumina, esekwe kulandelelwano nge-Synthesis (SBS), yinto entsha ye-NGS eyamkelwe kwihlabathi jikelele, enoxanduva lokuvelisa ngaphezulu kwe-90% yedatha yolandelelwano lwehlabathi.Umthetho-siseko we-SBS ubandakanya umboniso we-fluorescent obhalwe izitermina ezibuyiselwa umva njengoko i-dNTP nganye yongezwa, kwaye emva koko yacandwa ukuvumela ukufakwa kwesiseko esilandelayo.Ngazo zone ii-dNTP ezine-terminator-bound bounded dNTP ezikhoyo kumjikelo ngamnye wolandelelwano, ukhuphiswano lwendalo lunciphisa ukubandakanywa kokubandakanya.Ubuchwephesha obuguquguqukayo buxhasa amathala eencwadi afundwe kunye kunye nesiphelo sesibini, silungiselela uluhlu lwezicelo ze-genomic.I-Illumina yokulandelanisa i-high-throughput i-high-throughput ibeka njengelitye lembombo kuphando lwe-genomics, ixhobisa izazinzulu ukuba ziveze ubucukubhede be-genomes ngeenkcukacha ezingenakulinganiswa kunye nokusebenza kakuhle.

I-DNBSEQ, ephuhliswe yi-BGI, yenye itekhnoloji ye-NGS ekwaziyo ukuhla ngakumbi kwiindleko zolandelelwano kunye nokunyusa umthamo.Ukulungiswa kweelayibrari ze-DNBSEQ kubandakanya ukuhlukana kwe-DNA, ukulungiswa kwe-ssDNA kunye nokukhulisa isangqa esijikelezayo ukufumana i-nanoballs ye-DNA (DNB).Ezi ke ziye zilayishwe kumphezulu oqinileyo kwaye emva koko zilandelelaniswe ngokudityaniswa kweProbe-Anchor Synthesis (cPAS).

Inkonzo yethu yolandelelwano yethala leencwadi eyenziwe kwangaphambili iququzelela abathengi ekulungiseleleni iilayibrari zabo ezilandelelanayo ezivela kwimithombo eyahlukeneyo (mRNA, genome yonke, amplicon, phakathi kwabanye).Emva koko, la mathala eencwadi anokuthunyelwa kumaziko ethu okulandelelanisa ulawulo lomgangatho kunye nokulandelelana kwiiplatifti ze-Illumina okanye ze-BGI.

Qhagamshelana nathi

Iinkcukacha zeNkonzo

Iziphumo zeDemo

Iimbonakalo

●Amaqonga:Illumina NovaSeq 6000, NovaSeq , HiSeq X Ten kunye ne-BGI-DNB-T7

●Iimowudi zolandelelwano:PE50, PE100, PE150, PE250

●Ukulawulwa komgangatho wamathala eencwadi phambi kolandelelwano

●Ukulandelelanisa ukuhanjiswa kwedatha kunye ne-QC:ukuhanjiswa kwengxelo ye-QC kunye nedatha ekrwada kwifomathi ye-fastq emva kwe-demultiplexing kunye nokucoca ukufundwa kwe-Q30.

Iinkonzo eziluncedo

●Ukuguquguquka kweenkonzo zolandelelwano:umthengi unokukhetha ukulandelelanisa ngelayini, iseli yokuhamba okanye isixa sedatha.

●Ukuguquguquka kwamaqonga:Amathala eencwadi e-DNB anokudluliselwa kwiiplatifti ze-Illumina

●Amava abanzi kwiqonga lokulandelelana kwe-Illumina:ngamawaka eeprojekthi ezivaliweyo ezineentlobo ezahlukeneyo.

●Ukuhanjiswa kolandelelwano lwengxelo ye-QC:kunye neemetrics ezisemgangathweni, ukuchaneka kwedatha kunye nokusebenza ngokubanzi kweprojekthi yokulandelelana.

●Inkqubo yolandelelwano lwabantu abadala:ngexesha elifutshane lokujika.

●Ulawulo loMgangatho olungqongqo: siphumeza iimfuno ezingqongqo ze-QC ukuqinisekisa ukuhanjiswa kweziphumo ezihlala zikumgangatho ophezulu.

IiMfuno zeSampulu*

Ulandelelwano lweNdlela engaphelelanga

Isixa seDatha (X)	Ugxininiso (qPCR/nM)	Umthamo
X ≤ 50 Gb	≥ 2 nM	≥ 20 μl
50 Gb ≤ X < 100Gb	≥ 3 nM	≥ 20 μl
X ≥ 100 Gb	≥ 4 nM	≥ 20 μl

Indlela enye (Illumina)

Iqonga	Ugxininiso (qPCR/nM)	Umthamo
HiSeq X Ten	≥ 2 nM	≥ 20 μl
NovaSeq 6000 SP	≥ 1 nM	≥ 25 μl
NovaSeq 6000 S4	≥ 1.5 nM	≥ 25 μl
NovaSeq X	≥ 1.5 nM	≥ 25 μl
I-BGI-DNBSEQ-T7	≥ 1.5 nM	≥ 25 μl

Ukongeza kugxininiso kunye nenani elipheleleyo, ipateni efanelekileyo yokuphakama nayo iyafuneka.

Nceda ufikelele kuthi ukuba iisampulu zakho azifikeleli kwiimfuno zemathiriyeli zokuqalisa.

Qhagamshelana nathi

Ukuhanjiswa kwenkonzo

Ulawulo lomgangatho wethala leencwadi

Ukulandelelanisa

Ulawulo lomgangatho wedatha

Unikezelo lweprojekthi

Ngaphambili: I-DNA / i-RNA yokulandelelana -I-PacBio Sequencer

Okulandelayo: I-DNA / RNA Ukulandelelana - i-Nanopore Sequencer

Ingxelo yeThala leeNcwadi le-QC

Ingxelo engomgangatho wethala leencwadi inikwa phambi kolandelelwano, uvavanyo lwesixa-mali sethala leencwadi, nokwahlulwa.

Ukulandelelanisa ingxelo ye-QC

Itheyibhile 1. Iinkcukacha-manani ngokulandelelana kwedatha.

Isampulu yesazisi	BMKID	Ufunda ekrwada	Idatha ekrwada (bp)	Ukufundwa okucocekileyo (%)	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