Awọn ọja

DNA/RNA lesese -PacBio Sequencer

Syeed ipasẹ PacBio jẹ ipilẹ itẹlera kika gigun, eyiti o tun mọ bi ọkan ninu awọn imọ-ẹrọ Sequencing-Iran Kẹta (TGS).Imọ-ẹrọ mojuto, akoko gidi-molecule nikan (SMRT), n fun irandiran ti kika pẹlu mewa ti kilo-mimọ ni ipari.Lori ipilẹ ti “Sequencing-by-Synthesis”, ipinnu nucleotide ẹyọkan jẹ aṣeyọri nipasẹ ipo igbi Zero-mode(ZMW), nibiti iwọn didun ti o lopin nikan ni isalẹ (ojula ti iṣelọpọ moleku), ti tan imọlẹ.Ni afikun, tito lẹsẹsẹ SMRT yago fun aibikita ọkọọkan-pato ninu eto NGS, ni pe pupọ julọ awọn igbesẹ imudara PCR ko nilo ni ilana ikole ile-ikawe.

Platform: Atẹle II, Revio

Pe wa

Awọn alaye Iṣẹ

Awọn abajade Ririnkiri

Awọn ẹya ara ẹrọ

Awọn ipo itọsẹ meji lori atẹle PacBio: Kika Gigun Tesiwaju (CLR) ati Ipinnu Ijẹwọgba Ipin (CCS)

Ipo Atẹle

Iwon ikawe

Data o tumq siIkore (Ni ẹẹẹkeji)

Nikan-mimọYiye

Awọn ohun elo

CLR

20Kb, 30Kb, ati bẹbẹ lọ.

80 Gb si 130 GB

Isunmọ.85%

De novo, SV pipe, ati be be lo.

CCS

15-20 Kb

14 si 40 Gb/Sẹẹli (Atẹle II)

70 si 110 Gb/Sẹli (Revio)

Da lori awọn ayẹwo

Isunmọ.99%

De novo, SNP/Indel/SV pipe, Iso-Seq,

Lafiwe ti iṣẹ ati awọn ẹya ara ẹrọ ti Revio ati Sequel II

Awọn ofin	Atele II eto	Revio eto	Alekun
Ti o ga iwuwo	8 million ZMWs	25 million ZMWs	3x
Awọn ipele ominira	1	4	4x
Awọn akoko ṣiṣe kukuru	30 wakati	24 wakati	1.25x
30X HiFi eda eniyan genomes / odun	88	1.300	15x lapapọ

Awọn anfani Iṣẹ

● Lori 8 years iriri lori PacBio sequencing Syeed pẹlu egbegberun ti titi ise agbese pẹlu orisirisi eya.

● Ni ipese ni kikun pẹlu awọn iru ẹrọ PacBio Sequencing tuntun, Revio lati ṣe iṣeduro iṣelọpọ ilana ilana to to.

● Yiyara akoko titan-yika, ikore data ti o ga julọ ati data deede diẹ sii.

● Ti ṣe alabapin ninu awọn ọgọọgọrun awọn atẹjade ti o da lori ipa-giga ti PacBio.

Awọn ibeere apẹẹrẹ

Apeere Iru

Iye

Ifojusi (Qubit®)

Iwọn didun

Mimo

Awọn miiran

DNA jinomiki

Da lori Data ibeere

≥50 ng/μl

≥15μl

OD260/280 = 1.7-2.2;
OD260/230=1.8-2.5;
Ko tente oke ni 260 nmko si kontaminesonu

Ifojusi nilo lati ṣe iwọn nipasẹ Qubit ati Qubit/Nanopore = 0.8-2.5

Apapọ RNA

≥1.2μg

≥120 ng/μl

≥15μl

OD260 / 280 = 1.7-2.5;
OD260/230=0.5-2.5;ko si kontaminesonu

RIN iye ≥7.5

5≥28S/18S≥1

Ṣiṣẹ Iṣẹ Iṣẹ

Apeere igbaradi

Ikole ìkàwé

Titele

Itupalẹ data

Ifijiṣẹ ise agbese

Ti tẹlẹ: OEM olupese Mrna Sequencing Service - Long ti kii-ifaminsi lesese-Illumina – Biomarker

Itele: Illumina ati BGI

1.In-house Data Ikore

Data ti ipilẹṣẹ lati awọn sẹẹli 63 CCS (lati awọn ẹya 26)

DATA-PacBio-CCS-15 Kb	Apapọ	O pọju	Min	Agbedemeji
Ikore - awọn ikawe (Gb)	421.12	544.27	221.38	426.58
Yiled - 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
Apapọ ipari-Polymerase	67,995	89.379	49.664	66.433
Apapọ ipari-Subreads	15.866	21.036	11.657	16.012
Apapọ ipari-CCS	14.489	19.074	8.575	14.655

Data ti ipilẹṣẹ lati awọn sẹẹli 16 CLR (lati awọn ẹya 76)

DATA-PacBio-CLR-30Kb	Apapọ	O pọju	Min	Agbedemeji
Ikore - awọn ikawe (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
Apapọ ipari-Polymerase	22.063	48.886	8.747	21.555
Apapọ ipari-Subreads	17.720	27.225	8.293	17.779

2.Data QC - RirinkiriAwọn iṣiro lori ikore data

Apeere	ccs Say Nọm	Lapapọ Awọn ipilẹ ccs (bp)	ccs Ka N50 (bp)	Itumọ Gigun (bp)	ccs kika ti o gun julọ (bp)	Awọn ipilẹ abẹlẹ (bp)	oṣuwọn ccs (%)
PB_BMKxxx	3,444,159	54.164.122.586	15.728	15.726	36,110	863,326,330,465	6.27