Nā huahana

Ka hoʻokaʻina DNA/RNA -PacBio Sequencer

DNA/RNA sequencing -PacBio Sequencer Hōʻike Kiʻi

ʻO PacBio sequencing platform he papa kuhikuhi lōʻihi heluhelu ʻia, ʻike ʻia ʻo ia kekahi o nā ʻenehana Sequencing Third-Generation Sequencing (TGS).ʻO ka ʻenehana koʻikoʻi, hoʻokahi-molele manawa maoli (SMRT), hoʻoikaika i ka hana o nā heluhelu me nā ʻumi kilo-base ka lōʻihi.Ma ke kumu o "Sequencing-by-Synthesis", ua hoʻokō ʻia ka hoʻonā nucleotide hoʻokahi e Zero-mode waveguide (ZMW), kahi i hoʻomālamalama ʻia ai ka leo liʻiliʻi ma lalo (kahi o ka mole synthesis).Eia hou, pale ka SMRT sequencing nui i ka sequence-specific bias in NGS system, no ka mea, ʻaʻole koi ʻia ka hapa nui o nā ʻanuʻu PCR amplification ma ke kaʻina hana hale waihona puke.

Papahana: Sequel II, Revio

Kāhea iā mā˚ou

Nā kikoʻī lawelawe

Nā hopena hōʻike

Nā hiʻohiʻona

ʻElua mau ʻano hoʻohālikelike ma ka Sequencer PacBio: Hoʻomau i ka heluhelu lōʻihi (CLR) a me Circular Consensus Read (CCS)

Ke ʻano Sequencing

Nui o ka waihona

ʻIke ʻIke ManaʻoHua mai (no ke kelepona)

kahua hoʻokahiKa pololei

Nā noi

CLR

20Kb, 30Kb, etc.

80 Gb i 130 Gb

Ma kahi o.85%

De novo, kelepona SV, etc.

CCS

15-20 Kb

14 a i 40 Gb/Cell (Sequel II)

70 a hiki i 110 Gb/Cele (Revio)

Pili i nā laʻana

Ma kahi o.99%

De novo, Kāhea SNP/Indel/SV, Iso-Seq,

Hoʻohālikelike i ka hana a me nā hiʻohiʻona o Revio a me Sequel II

ʻŌlelo	Pūnaehana Sequel II	Pūnaehana Revio	Hoʻonui
Kiʻekiʻe kiʻekiʻe	8 miliona ZMW	25 miliona ZMW	3x
Nā pae kūʻokoʻa	1	4	4x
Nā manawa holo pōkole	30 hola	24 hola	1.25x
30X HiFi kanaka genome / makahiki	88	1,300	15x ka nui

Pono lawelawe

● Ma luna o 8 mau makahiki ʻike ma PacBio sequencing platform me nā kaukani o nā papahana pani me nā ʻano ʻano like ʻole.

● Hoʻolako piha ʻia me nā kahua PacBio Sequencing hou loa, ʻo Revio e hōʻoiaʻiʻo i ka lawa o ka sequencing throughput.

● ʻOi aku ka wikiwiki o ka manawa huli, ʻoi aku ka nui o ka ʻikepili a me ka ʻikepili pololei.

● Hāʻawi ʻia i loko o nā haneli o nā puke paʻi kiʻekiʻe ʻo PacBio.

Nā Koina Laʻana

ʻAno Laʻana

Ka nui

Hoʻopaʻa （Qubit®）

Volume

Maemae

ʻO nā mea ʻē aʻe

DNA Genomic

E hilinaʻi i ka ʻikepili koi

≥50 ng/μl

≥15μl

OD260/280=1.7-2.2;
OD260/230=1.8-2.5;
Maikaʻi ka piko ma 260 nm，ʻaʻohe haumia

Pono e ana ʻia ka nui e Qubit a me Qubit/Nanopore = 0.8-2.5

Huina RNA

≥1.2μg

≥120 ng/μl

≥15μl

OD260/280=1.7-2.5;
OD260/230=0.5-2.5;ʻaʻohe haumia

Waiwai RIN ≥7.5

5≥28S/18S≥1

Hana Hana Hana

Hoʻomākaukau laʻana

Kūkulu hale waihona puke

Ke kaʻina ʻana

ʻIkepili ʻikepili

Hāʻawi papahana

Mua: Mea hana OEM Mrna Sequencing Service - Long non-coding sequencing-Illumina - Biomarker

Aʻe: Illumina a me BGI

1.Ma loko o ka hale ʻikepili hua

ʻIke i hana ʻia mai 63 CCS cell (mai 26 ʻano)

ʻIkepili-PacBio-CCS-15 Kb	Awelika	Max	Min	Mediana
Hāʻawi ʻia - nā heluhelu liʻiliʻi (Gb)	421.12	544.27	221.38	426.58
Hāʻawi ʻia - CCS(Gb)	25.93	38.59	10.86	25.43
Polymerase N50	145,651	175,430	118,118	144,689
Heluhelu N50	17,509	23,924	12,485	17,584
CCS N50	14,490	19,034	9,876	14,747
Ka lōʻihi-Polymerase	67,995	89,379	49,664	66,433
Awelika lōʻihi-subreads	15,866	21,036	11,657	16,012
Awelika lōʻihi-CCS	14,489	19,074	8,575	14,655

Nā ʻikepili i hana ʻia mai nā pūnaewele CLR 16 (mai 76 mau ʻano)

DATA-PacBio-CLR-30Kb	Awelika	Max	Min	Mediana
Hāʻawi ʻia - nā heluhelu liʻiliʻi (Gb)	142.20	291.40	50.55	142.49
Polymerase N50	39,456	121,191	15,389	35,231
Heluhelu N50	28,490	41,012	14,430	29,063
Ka lōʻihi-Polymerase	22,063	48,886	8,747	21,555
Awelika lōʻihi-subreads	17,720	27,225	8,293	17,779

2.ʻIkepili QC - DemoHeluhelu e pili ana i ka loaʻa ʻikepili

Laʻana	ccs Heluhelu Num	Huina ccs kumu (bp)	ccs Heluhelu N50 (bp)	ccs Mean Length (bp)	ccs Heluhelu lōʻihi loa (bp)	Heluhelu hou Nā kumu (bp)	helu ccs(%)
PB_BMKxxx	3,444,159	54,164,122,586	15,728	15,726	36,110	863,326,330,465	6.27