Imikhiqizo

Ukulandelana kwe-DNA/RNA – I-Nanopore Sequencer

Ukulandelana kwe-DNA/RNA – Isithombe Esifakiwe se-Nanopore Sequencer

Ukulandelana kwe-ONT kuwubuchwepheshe bokulandelana kwesignali kagesi yesikhathi sangempela obusekelwe kuma-nanopores, isimiso sokulandelana kwepulatifomu ngayinye siyafana. I-DNA/RNA enezintambo ezimbili izobopha kuphrotheni enezimbobo ezincane efakwe ku-biofilm futhi ikhululeke ngaphansi kokuhola kwephrotheni yemoto, ngaphansi kwesenzo somehluko we-voltage kuzo zombili izinhlangothi ze-biofilm, izintambo ze-DNA/RNA zidlula kuphrotheni yesiteshi se-nanopore ngesivinini esithile. Ngenxa yokwehluka kwezakhiwo zamakhemikhali zezisekelo ezahlukene ku-strand ye-DNA/RNA, lapho isisekelo esisodwa noma i-molecule ye-DNA idlula ku-channel ye-nanopore, kuzobangela ushintsho lwezimpawu zikagesi ezahlukene. Ngokuthola nokuhambisana nalezi zimpawu, izinhlobo zesisekelo ezihambisanayo zingabalwa, futhi ukutholwa kwesikhathi sangempela kokulandelana kungaqedwa.

Xhumana nathi

Imininingwane Yesevisi

Umphumela Wedemo

Izici Zemininingwane Yesevisi

Ipulatifomu	Usayizi Welabhulali	Ukukhiqizwa Kwedatha Yethiyori (Ngeseli Ngayinye)	Ukunemba Okuyisisekelo Esisodwa	Izicelo
I-Nanopore	8Kb, 10kb, 20kb, Ultralong, cDNA-PCR	70-90Gb/Iseli	85-92%	Ukubizwa kwe-SV, i-De novo, Ukulandelana kobude obugcwele, i-Iso-Seq, isichasiselo se-Gene, Ukutholwa kwe-methylation ye-DNA

Izinzuzo Zesevisi

● Isipiliyoni seminyaka engaphezu kwemi-5 ku-PacBio sequencing platform enezinkulungwane zamaphrojekthi avaliwe anezinhlobo ezahlukene.
● I-BMKGENE ingumlingani osemthethweni we-Oxford Nanopore, enesitifiketi se-RNA/DNA esiphindwe kabili.
● Kunezinhlobo ezijwayelekile zama-sequencers anemishini ephelele kanye nomphumela owanele wokulandelana.
● Ngokusekelwe epulatifomu ye-Nanopore, ucwaningo lwezilwane nezitshalo ezingaphezu kwe-10 lwe-Denovo luye lwanyatheliswa emaphephandabeni adumile emhlabeni jikelele.

Izidingo Zesampula

Uhlobo Lwesampula

Inani

Ukugxila (Qubit ®)

Ivolumu

Ubumsulwa

Abanye

I-DNA ye-Genomic

Thembela Esidingweni Sedatha

≥20ng/μl

≥15μl

OD260/280=1.7-2.2;

OD260/230≥1.5；

Sula ukuphakama ku-260 nm, akukho ukungcola

Ukuhlushwa kudinga ukukalwa nge-Qubit kanye ne-Qubit/Nanopore ≤ 2

I-RNA ephelele

≥1.2μg

≥100μg/μl

≥15μl

OD260/280=1.7-2.5;

OD260/230=0.5-2.5；akukho ukungcola

Inani le-RIN ≥7.5

Ukuhamba Komsebenzi Wesevisi

Ukulungiselela isampula

Ukwakhiwa komtapo wolwazi

Ukulandelana

Ukuhlaziywa kwedatha

Ukulethwa kwephrojekthi

Okwedlule: Imitapo yolwazi eyenziwe kusengaphambili ye-Illumina

Olandelayo: Ukulandelana Kwezingcezu Ezihlanganisiwe Zendawo Ethile (SLAF-Seq)

Ukuhlolwa Kwekhwalithi Yedatha Yesampula Ye-DNA

Ithebula 1. Izibalo ngedatha ehlanzekile.

I-BMKID	i-rawSeqNum	i-rawSumBase	i-cleanSeqNum	i-cleanSumBase	i-cleanN50Len	i-cleanN90Len	i-cleanMeanLen	i-cleanMaxLen	i-cleanMeanQual
I-DNA_BMK01	1,218,239	26.37	1,121,736	25.90	28,014	15,764	23,090	143,181	9