Ubushinwa ADN / RNA ikurikirana -PacBio Uruganda rukora n'ababikora |BMK

Urutonde rwa ADN / RNA -PacBio Ikurikirana

DNA/RNA sequencing -PacBio Sequencer Featured Image

Urubuga rwa PacBio rukurikirana ni urubuga rwasomwe igihe kirekire, ruzwi kandi nka bumwe mu buhanga bwa gatatu (TGS).Tekinoroji yibanze, molekile imwe-nyayo-nyayo (SMRT), iha imbaraga ibisekuruza byasomwe hamwe na kilo-shingiro muburebure.Hishimikijwe "Gukurikirana-by-Synthesis", gukemura nucleotide imwe bigerwaho na Zero-mode waveguide (ZMW), aho ijwi rito gusa hepfo (site ya synthesis ya molekile), rimurikirwa.Mubyongeyeho, urutonde rwa SMRT rwirinda ahanini kubogama kubogamye muri sisitemu ya NGS, kuberako intambwe nyinshi zo kongera PCR zidasabwa mubikorwa byo kubaka isomero.

Twandikire

Ibisobanuro bya serivisi

Ibisubizo bya Demo

Ibiranga

Uburyo bubiri bukurikirana kuri PacBio ikurikiranye: Gukomeza Birebire Gusoma (CLR) hamwe no Kuzenguruka Byasomwe (CCS)

Uburyo bukurikirana	Ingano y'Isomero	IbyatanzweTanga umusaruro (Akagari)	Urufatiro-shingiroUkuri	Porogaramu
CLR	20Kb, 30Kb, n'ibindi.	80 Gb kugeza 130 Gb	Hafi.85%	De novo, SV guhamagara, nibindi
CCS	15-20 Kb	14 kugeza 40 Gb / selile (CCS)	Hafi.99%	De novo, SNP / Indel / SV guhamagara, Iso-Seq,

Inyungu za serivisi

ØUburambe bwimyaka 7 kurubuga rwa PacBio hamwe nibikorwa ibihumbi n'ibihumbi bifunze hamwe nubwoko butandukanye.

ØByuzuye byuzuye bya platform ya PacBio ikurikirana, Sequel II kugirango yizere ko byinjira neza

ØByihuta Byihuta, umusaruro mwinshi hamwe namakuru yukuri.

ØYatanze umusanzu mu majana-yingirakamaro cyane ya PacBio

Icyitegererezo

Ubwoko bw'icyitegererezo	Umubare	Kwibanda （Qubit®）	Umubumbe	Isuku	Abandi
ADN ya ADN	Biterwa nibisabwa	≥20 ng / μl	≥15μl	OD260 / 280 = 1.6-2.3; OD260 / 230 = 1.5-2.5 ； Kuraho impinga kuri 260 nm ，nta byanduye	Kwishyira hamwe bigomba gupimwa na Qubit na Qubit / Nanopore = 0.8-2.0
RNA Yuzuye	.51.5μg	≥40 ng / μl	≥15μl	OD260 / 280 = 1.7-2.5; OD260 / 230 = 0.5-2.5 ；nta byanduye	RIN agaciro ≥7.5

Ibikorwa bya serivisi

Icyitegererezo

Kubaka isomero

Urukurikirane

Isesengura ryamakuru

Gutanga umushinga

Mbere: Isesengura ryinshi rya Segregant

Ibikurikira: Kugereranya Genomika

1.Mu nzu Data Yitanga

Amakuru yatanzwe muri selile 63 CCS (kuva mumoko 26)

Data-Pacbio-CCS-15 Kb	Impuzandengo	Icyiza	Min	Median
Umusaruro - subreads (Gb)	421.12	544.27	221.38	426.58
Yile - CCS (Gb)	25.93	38.59	10.86	25.43
Polymerase N50	145,651	175.430	118.118	144,689
Subreads N50	17,509	23,924	12.485	17.584
CCS N50	14.490	19.034	9.876	14.747
Impuzandengo y'uburebure-Polymerase	67,995	89.379	49,664	66.433
Impuzandengo y'uburebure-Gusoma	15.866	21.036	11,657	16,012
Impuzandengo y'uburebure-CCS	14.489	19.074	8.575	14,655

Amakuru yatanzwe muri selile 16 za CLR (kuva mumoko 76)

DATA-Pacbio-CLR-30Kb	Impuzandengo	Icyiza	Min	Median
Umusaruro - subreads (Gb)	142.20	291.40	50.55	142.49
Polymerase N50	39,456	121.191	15.389	35,231
Subreads N50	28.490	41,012	14.430	29,063
Impuzandengo y'uburebure-Polymerase	22.063	48.886	8.747	21.555
Impuzandengo y'uburebure-Gusoma	17.720	27.225	8.293	17,779

2.Data QC - DemoImibare ku musaruro wamakuru

Icyitegererezo	ccs Isoma Umubare	Ccs zose zifatizo (bp)	ccs Isoma N50 (bp)	ccs Bisobanura Uburebure (bp)	ccs Gusoma Birebire (bp)	Subreads Base (bp)	igipimo cya ccs (%)
PB_BMKxxx	3,444.159	54.164.122.586	15,728	15.726	36.110	863.326.330.465	6.27