Tsarin DNA/RNA -PacBio Sequencer

Sequencing DNA/RNA -PacBio Sequencer Featured Image

PacBio dandali ne na jerin abubuwan da aka daɗe ana karantawa, wanda kuma aka sani da ɗayan fasahar Sequencing na ƙarni na uku (TGS).Babban fasahar, ainihin-kwale-kwale-kwale-kwale-kwale-kwale (SMRT), yana ba da ƙarfin ƙarni na karantawa tare da dubun-kwanan kilogiram cikin tsayi.A kan tushen "Sequencing-by-Synthesis", ƙudurin nucleotide guda ɗaya yana samuwa ta hanyar Zero-mode waveguide (ZMW), inda aka haskaka ƙayyadaddun ƙarar ƙarar kawai a ƙasa (shafin haɗin ƙwayoyin ƙwayoyin cuta).Bugu da ƙari, jeri na SMRT ya fi guje wa ƙayyadaddun ƙayyadaddun jeri a cikin tsarin NGS, a cikin cewa yawancin matakan haɓaka PCR ba a buƙatar su a tsarin ginin ɗakin karatu.

Platform: Mabiyi II, Revio

Tuntube Mu

Cikakkun Sabis

Sakamakon Demo

Siffofin

Hanyoyin jeri guda biyu akan mabiyi na PacBio: Ci gaba da Karatu (CLR) da Karatun Ijma'i (CCS)

Yanayin Jeri

Girman Laburare

Bayanan Ka'idarHaihuwa (Kowace Tantanin halitta)

Single-tusheDaidaito

Aikace-aikace

CLR

20Kb, 30kb, da dai sauransu.

Daga 80 zuwa 130 GBp

Kimanin85%

Da novo, kiran SV, da dai sauransu.

CCS

15-20 Kb

14 zuwa 40 Gb/cell (Sequel II)

70 zuwa 110 Gb/cell (Revio)

Ya dogara da samfurori

Kimanin99%

Da novo, SNP/Indel/SV kira, Iso-Seq,

Kwatanta ayyuka da fasalulluka na Revio da Sequel II

Sharuɗɗan	Mabiyi II tsarin	Tsarin Revio	Ƙara
Mafi girma yawa	8 miliyan ZMWs	ZMW miliyan 25	3x
Matakai masu zaman kansu	1	4	4x
Gajeren lokutan gudu	30 hours	awa 24	1.25x ku
30X HiFi genomes na mutum / shekara	88	1,300	15x gaba daya

Amfanin Sabis

● Fiye da shekaru 8 gwaninta akan dandamali na jerin abubuwan PacBio tare da dubban rufaffiyar ayyukan tare da nau'ikan nau'ikan daban-daban.

● Cikakken sanye take da sabbin dandamali na Sequencing na PacBio, Revio don ba da garantin isassun kayan sarrafawa.

● Saurin jujjuyawar lokaci, yawan amfanin bayanai da ƙarin cikakkun bayanai.

● An ba da gudummawa a ɗaruruwan wallafe-wallafen PacBio masu tasiri.

Samfuran Bukatun

Nau'in Misali

Adadin

Tattaunawa (Qubit®)

Ƙarar

Tsafta

Wasu

Genomic DNA

Ya dogara da Bukatun Bayanai

≥50 ng/μl

≥15 ml

OD260/280=1.7-2.2;
OD260/230=1.8-2.5;
Matsakaicin tsayi a 260 nmbabu gurbacewa

Ana buƙatar auna hankali ta hanyar Qubit da Qubit/Nanopore = 0.8-2.5

Jimlar RNA

≥1.2μg

≥120 ng/μl

≥15 ml

OD260/280=1.7-2.5;
OD260/230=0.5-2.5;babu gurbacewa

Darajar RIN ≥7.5

5≥28S/18S≥1

Gudun Aikin Sabis

Samfurin shiri

Gina ɗakin karatu

Jeri

Binciken bayanai

Isar da aikin

Na baya: Ma'aikatan OEM Mrna Sequencing Sabis - Dogon jerin abubuwan da ba su da lamba-Illumina - Biomarker

Na gaba: Sequencing DNA/RNA - Illumina Sequencer

1.In-gida Data Haihuwa

Bayanan da aka samar daga sel 63 CCS (daga nau'ikan 26)

DATA-PacBio-CCS-15 Kb	Matsakaicin	Max	Min	Matsakaici
Haɓaka - Rarraba (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
Mai Rarraba N50	17,509	23,924	12,485	17,584
CCS N50	14,490	19,034	9,876	14,747
Matsakaicin tsayi-Polymerase	67,995	89,379	49,664	66,433
Matsakaicin tsayi-Kayan abinci	15,866	21,036	11,657	16,012
Matsakaicin tsayi-CCS	14,489	19,074	8,575	14,655

Bayanan da aka samar daga sel 16 CLR (daga nau'ikan 76)

DATA-PacBio-CLR-30Kb	Matsakaicin	Max	Min	Matsakaici
Haɓaka - Rarraba (Gb)	142.20	291.40	50.55	142.49
Polymerase N50	39,456	121,191	15,389	35,231
Mai Rarraba N50	28,490	41,012	14,430	29,063
Matsakaicin tsayi-Polymerase	22,063	48,886	8,747	21,555
Matsakaicin tsayi-Kayan abinci	17,720	27,225	8,293	17,779

2.Data QC - DemoKididdigar kan yawan yawan bayanai

Misali	ccs Karanta Lambobi	Jimlar Bases na ccs (bp)	ccs yana karanta N50 (bp)	ccs Ma'ana Tsawon (bp)	ccs Mafi Dadewar Karatu (bp)	Rubutun ƙasa (bp)	ƙimar ccs (%)
PB_BMKxxx	3,444,159	54,164,122,586	15,728	15,726	36,110	863,326,330,465	6.27