Olink NextSeq 2000 Ongorora Sequencing System

Document note

Iyo Olink® Ongorora Mushandisi bhuku, doc nr 1153, haichashandi, uye yakatsiviwa nemagwaro anotevera:

• Olink® Ongorora Pamusoroview Mushandisi Manual, doc nr 1187
• Olink® Ongorora 384 User Manual, doc nr 1188
• Olink® Ongorora 4 x 384 User Manual, doc nr 1189
• Olink® Ongorora 1536 & Kuwedzera Mushandisi Manual, doc nr 1190
• Olink® Ongorora 3072 User Manual, doc nr 1191
• Olink® Ongorora Sequencing uchishandisa NextSeq 550 Mushandisi Manual, doc nr 1192
• Olink® Ongorora Sequencing uchishandisa NextSeq 2000 Mushandisi Manual, doc nr 1193
• Olink® Ongorora Sequencing uchishandisa NovaSeq 6000 User Manual, doc nr 1194

Nhanganyaya

Chinangwa chekushandisa

Olink® Ongorora ndeye multiplex immunoassay chikuva chekuwanikwa kweprotein biomarker yevanhu. Chigadzirwa chacho chakagadzirirwa Kutsvaga Kushandisa Chete, uye kwete kushandiswa mukuongorora maitiro. Basa remurabhoritari richaitwa chete nevashandi vakadzidziswa. Kugadziriswa kwedata kuchaitwa chete nevashandi vakadzidziswa. Migumisiro inofanirwa kushandiswa nevatsvakurudzi pamwe chete nemamwe makiriniki kana marabhoritari zvakawanikwa.

Nezve bhuku rino

Iri Bhuku reMushandisi rinopa mirairo inodiwa kutevedzana Olink® Ongorora Maraibhurari paIllumina® NextSeq™ 2000. Mirayiridzo inofanira kunyatsoteverwa uye zvakajeka. Chero kutsauka mumatanho ese emurabhoritari kunogona kukonzera kukanganisa data. Usati watanga kufambiswa kwerabhoritari, bvunza iyo Olink® Ongorora Pamusoroview Mushandisi Manual yekusuma papuratifomu, kusanganisira ruzivo nezve reagents, zvishandiso uye zvinyorwa zvinodiwa, pamusoroview yemafambiro ebasa, pamwe chete nemirayiridzo yerabhoritari. Kuti uwane mirairo yemaitirwo eOlink® Ongorora Reagent Kits, tarisa kune inoshanda Olink® Ongorora Mushandisi Manual. Kuti uwane data yekugadzirisa uye kuongororwa kweOlink® Ongorora kutevedzana mhedzisiro, tarisa iyo Olink® MyData Cloud User Guide. Ese maratidziro uye kodzero dzekodzero dziri muchinhu ichi zvinhu zveOlink® Proteomics AB, kunze kwekunge zvataurwa neimwe nzira.

Tsigiro yehunyanzvi

Kuti uwane rutsigiro rwehunyanzvi, bata Olink Proteomics pa: support@olink.com.

Laboratory mirayiridzo

Ichi chitsauko chinopa mirairo yekuteedzera Olink Libraries paNextSeq™ 2000 uchishandisa iyo NextSeq™ 1000/2000 P2 Reagents (100 Cycles) v3. Iyo protocol inoshandiswa pakutevedzana ndeyekugadziriswa kweIllumina® standard NGS workflow yeIllumina® NextSeq™ 2000. Usati waenderera mberi kutevedzana, iva nechokwadi chokuti hutano hweOlink Library yakacheneswa yakasimbiswa. Tarisa kune inoshanda Olink Ongorora Mushandisi Manual kune mirairo nezve kutonga kwemhando.

Ronga kutevedzana kumhanya

Imwe Olink Raibhurari inogona kutevedzana neNextSeq™ 2000 P2 inoyerera sero uye nekumhanya. Nhamba yeP2 inoyerera masero uye inomhanya inodiwa kuteedzera akasiyana Olink Ongorora Reagent Kits inotsanangurwa muTebhu 1. Kana kupfuura kumhanya kumwe kuchidiwa, dzokorora mirairo inotsanangurwa mubhuku rino.

Tafura 1. Sequencing run kuronga

Olink® Ongorora Reagent Kit	Nhamba yeOlink Libraries	Nhamba yemasero ekuyerera uye kumhanya (s)
Olink® Ongorora 384 Reagent Kit	1	1
Olink® Ongorora 4 x 384 Reagent Kit	4	4
Olink® Ongorora 1536 Reagent Kit	4	4
Olink® Ongorora Kuwedzera Reagent Kit	4	4
Olink® Ongorora 3072 Reagent Kit	8	8

Isa Olink® tsika yekubika

Sevha iyo Olink tsika resipi xml-file Olink_NSQ2K_P2_V1 mune yakakodzera chiridzwa folda.

CHERECHEDZA: Iyo Olink tsika yekubika inongoshanda neNextSeq™ 1000/2000 P2 Reagents (100 Cycles) v3 kit uye NextSeq™ 1000/2000 control software v1.2 kana v1.4.

Gadzirira sequencing reagents

Munguva iyi, iyo reagent cartridge ine clustering uye sequencing reagents inonyungudutswa uye sero rekuyerera rinogadzirira.

YAMBIRO: Iyo reagent cartridge ine makemikari anogona kuve ane njodzi. Pfeka midziyo yekudzivirira yakakwana uye urase mareagents akashandiswa zvinoenderana nezviyero zvinoshanda. Kuti uwane rumwe ruzivo, tarisa kuIllumina NextSeq 1000 uye 2000 System Guide (gwaro #1000000109376).

Gadzirira reagent cartridge

Kunyungudusa katuriji isina kuvhurwa inogona kuitwa uchishandisa nzira nhatu dzakasiyana: pakamuri tembiricha, mune inodzorwa yekugezesa mvura, kana mufiriji.

Gadzirira bhenji

• 1x NextSeq™ 1000/2000 P2 Reagent Cartridge (100 kutenderera)

Mirayiridzo

1. Chengetedza iyo reagent cartridge sezvakatsanangurwa muTafura 2.

Tafura 2. Reagent cartridge thawing nzira

Thawing nzira	Mirayiridzo
Pakamuri tembiricha	Pasina kuvhura bhegi resirivheri foil, isa katuriji yakaomeswa reagent pabhenji uye woinyungudutsa pamhepo tembiricha kwemaawa mapfumbamwe. Usapfuure maawa gumi nematanhatu. Ita shuwa kuti bhegi label rakatarisana kumusoro uye mweya iwoyo unogona kutenderera wakatenderedza cartridge.
Mukugeza nemvura	Pasina kuvhura bhegi resirivheri foil, isa katuriji yakaomeswa nechando yakanyura mumvura inodzorwa 25 °C yekugezera yemvura uye woirega ichinyunguduka kwemaawa matanhatu. Usapfuure maawa masere. Ita shuwa kuti bhegi label rakatarisana kumusoro uye kuti cartridge haitendeuke panguva yekunyunguduka. Dry cartridge zvakakwana nemucheka wepepa.
Mufiriji	Pasina kuvhura bhegi resirivheri foil, isa katuriji pabhenji uye uinyungudire pamhepo tembiricha kwemaawa matanhatu. Ita shuwa kuti bhegi label rakatarisana kumusoro uye mweya iwoyo unogona kutenderera wakatenderedza cartridge. Pedzisa kunyungudutsa cartridge mufiriji kwemaawa gumi nemaviri. Usapfuure maawa 12. Usati wamhanya, bvisa katuriji yakanyungudutswa isina kuvhurika kubva mufiriji uye uibvumire kuti isvike tembiricha yekamuri mukati memaminitsi gumi nemashanu, asi isingasviki 15 awa.

CHERECHEDZA: Makaturiji akanyungudutswa haagone kuomeswa nechando uye anofanira kuchengetwa pa4 ° C, kwenguva yakareba yemaawa makumi manomwe nemaviri.

Gadzirira kuyerera sero

Gadzirira bhenji

• 1x NextSeq™ 1000/2000 P2 Flow Cell

Mirayiridzo

1. Uya nefiriji inoyerera sero kune tembiricha yekamuri kwemaminetsi gumi-10.

Gadzirira Olink® Library yekutevedzana

Pakati peichi nhanho, iyo yakacheneswa uye yemhando inodzorwa Olink Raibhurari inonatswa kune yekupedzisira kurodha yekumisikidza. Ziva kuti Library denaturation inoitwa otomatiki pane chiridzwa.

Gadzirira bhenji

• Lib Tube, yakagadzirirwa zvinoenderana neiyo inoshanda Olink Ongorora Mushandisi Manual
• 1x RSB ine Pakati pe20
• MilliQ mvura
• 2x Microcentrifuge tubes (1.5 mL)
• Manyore pipette (10, 100 uye 1000 μL)
• Sefa mazano epipette

Usati watanga

• Nyora iyo Lib Tube kana yakaoma.
• Nyora iyo RSB yakaomeswa nechando neTween 20 pane tembiricha yemaminetsi gumi. Chengetedza pa +10 °C kusvikira washandiswa.
• Maka iwo maviri matsva 1.5 mL microcentrifuge chubhu sezvinotevera:
  • Maka imwe chubhu "Dil" (yeiyo 1:100 diluted Library)
  • Maka imwe chubhu "Seq" (yekugadzirira kurodha Raibhurari)

Mirayiridzo

1. Wedzera 495 ​​μL yeMilliQ mvura kune Dil Tube.
2. Vortex iyo Lib Tube uye itenderedze pasi muchidimbu.
3. Pamaoko endesa 5 μL kubva kuLib Tube kuenda kuDil Tube.
4. Vortex iyo Dil Tube uye itenderedze pasi muchidimbu.
5. Wedzera 20 μL yeRBS ine Pakati pemakumi maviri kuSeq Tube.
6. Nemaoko endesa makumi maviri μL kubva kuDil Tube kuenda kuSeq Tube.
7. Vortex iyo Seq Tube uye uitenderedze pasi muchidimbu.
8. Pakarepo enderera kune 2.5 Load kuyerera sero uye Olink® Library mune reagent cartridge.

CHERECHEDZA: Chengetedza Lib Tube (s) pa -20 °C kana paine zvingangoita kudzokorora (s).

Rodha kuyerera sero uye Olink® Library mune reagent cartridge

Munguva iyi, iyo sero yekuyerera uye yakanyungudutswa Olink Raibhurari inoiswa muine thawed reagent cartridge.

Gadzirira bhenji

• 1x yakanyungudutswa NextSeq™ 1000/2000 P2 Reagent Cartridge (100 cycles), yakagadzirirwa munhanho yapfuura.
• 1x NextSeq™ 1000/2000 P2 Flow Cell, yakagadzirirwa munhanho yapfuura
• Seq Tube (ine yakagadzirira kurodha yakanyungudutswa Olink Library), yakagadzirirwa mune yapfuura nhanho
• Chinyorwa pipette (100 μL)
• Pipette tip (1 mL)

Gadzirira cartridge

1. Bvisa cartridge kubva muhomwe yesirivha foil.
2. Pindura katuriji kagumi kuti unyatso kusanganisa ma reagents akanyungudika mukati.

CHERECHEDZA: Zvakajairika kunzwa zvinhu zvemukati zvichirira.

Isa sero yekuyerera mu cartridge

1. Kana wakagadzirira kurodha sero yekuyerera mu cartridge, bvisa iyo inoyerera sero kubva pasuru. Bata sero rekuyerera negrey tebhu, ine zita riri patebhu rakatarisa kumusoro. Shandisa magurovhosi matsva ehupfu kudzivirira kusvibisa girazi repamusoro pesero rekuyerera.
2. Isai iyo inoyerera sero mukati inoyerera sero slot pamberi pekaturiji. Kudzvanya kunonzwika kunoratidza kuti sero rekuyerera rakaiswa nemazvo.
3. Bvisa grey tab nekuidhonza kunze.

Isa iyo Olink® Library mu cartridge

1. Boora dura reLibhurari nechakachena 1 mL pipette tip.
2. Rodha 20 μL yeOlink Library kubva kuSeq Tube ichipinda muzasi medura reraibhurari.

Ita Olink® sequencing run

Munguva iyi, iyo Buffer cartridge ine yakatakurwa kuyerera sero uye Olink Library inoiswa muNextSeq™ 2000, uye kutevedzana kumhanya kunotanga uchishandisa Olink resipi.

Gadzirira bhenji

• 1x NextSeq™ 1000/2000 P2 Reagent Cartridge (matenderedzwa zana) akaremerwa neNextSeq™ 100/1000 P2000 Flow Cell uye yakanyungudutswa Olink Library, yakagadzirirwa munhanho yapfuura.

Gadzirisa iyo Run Mode

1. Kubva pane control software menyu, sarudza Settings.
2. Pasi peBaseSpace Sequence Hub Services & Proactive Support, sarudza Local Run Setup.
3. Sarudza Proactive Support Chete semamwe marongero. Iri basa rinoda kubatana kweinternet.
4. Sarudza iyo Hosting Nzvimbo yedata rako. Nzvimbo yekutambira inofanirwa kunge iri mukati kana padyo nenzvimbo yako.
5. Seta iyo Output Folder nzvimbo yeyazvino mhanyisa data raw. Sarudza Sarudza kufamba uye sarudza iyo inobuda folda.
6. Sarudza iyo Denature uye Dilute PaBhodhi cheki bhokisi kuti utore otomatiki uye uderedze Raibhurari iri pabhodhi chiridzwa.
7. Sarudza iyo Purge Reagent Cartridge cheki bhokisi kuti uzvichenese otomatiki reagents kune yakashandiswa reagents compartment ye cartridge.
8. Sarudza iyo Autocheck yesoftware inogadziridza bhokisi kuti utarise otomatiki zvigadziriso zvesoftware (sarudzo). Iri basa rinoda kubatana kweinternet.
9. Sarudza Save.

Rongedza run parameters

CHERECHEDZA: Murairo uyu unoshanda kune vhezheni 1.4 yeNextSeq™ 1000/2000 control software. Mamwe ematanho anotsanangurwa pazasi anogona kunge akasiyana kana uchishandisa shanduro v1.2

1. Kubva pane control software menyu, sarudza Tanga.
2. Sarudza Nemaoko Seta New Run uye tinya Setup.
3. Mune iyo Run Setup peji, gadzira iyo inomhanya paramita sezvinotevera:
   • MuRun Name ndima, isa yakasarudzika ID yekuyedza.
   • MuChinyorwa chekudonhedza Type Type, sarudza sarudzo yeKuverenga Imwe.
   • Isa nhamba yema cycles sezvinotevera:
     • Verenga 1: 24
     • Index 1: 0
     • Index 2: 0
     • Verenga 2: 0

CHERECHEDZA: Izvo zvakakosha kuti Read 1 yakaiswa ku24, zvikasadaro kumhanya kwese kuchakundikana. Rega mameseji ekunyevera paunenge uchipinda nhamba yematenderedzwa.

• MuCustom Primer Wells yekudonha-pasi rondedzero, sarudza Nha.
• Muchikamu cheCustom Recipe (optional), sarudza Sarudza kufamba uye sarudza tsika yekubika XML file Olink_NSQ2K_P2_V1. Sarudza Vhura.
• Usatore kunze Sample Sheet.
• Ita shuwa kuti iyo Output Folder nzvimbo ndeyechokwadi. Zvikasadaro, sarudza Sarudza kufamba uye sarudza yaunoda inobuda folda nzvimbo.
• MuDenature uye Dilute Onboard field, sarudza Inogoneswa kubva pane yekudonha-pasi runyorwa.
• Sarudza Prep.

Takura cartridge yakatakurwa

1. Sarudza Mutoro. Iyo visor yemidziyo inovhura uye tireyi inoburitswa.
2. Isa cartridge yakatakura patireyi ine label yakatarisa kumusoro uye kuyerera sero mukati mechiridzwa.
3. Sarudza Kuvhara.
4. Kana iyo cartridge yakanyatso kurodha, simbisa iyo yekumhanya paramita uye sarudza Sequence. Chiridzwa chinoita pre-run cheki yechiridzwa uye fluidics.
   • CHERECHEDZA: Munguva yekutarisa kwemvura, zvinotarisirwa kunzwa akati wandei achibuda.
5. Ita shuwa kuti kumhanya kunotanga mushure mekunge otomatiki pre-run cheki yapera (~ maminitsi gumi nemashanu). Iyo yekutevedzana yekumhanya nguva ingangoita 15h10 min.
   • CHERECHEDZA: Kune chero pre-run cheki kukanganisa, tarisa kune mirairo yemugadziri. Chenjerera kuti usarovera mukati kana kukanganisa iyo NextSeq™ 2000 panguva yekutevedzana kumhanya. Chiridzwa chacho chinonzwa nekudedera.
6. Chenesa nzvimbo yebasa.

Monitor Mhanyai Kufambira mberi

Olink inoshandisa NGS sekuverenga kuverenga kuti iverenge huwandu hwekutevedzana kunozivikanwa kuitira kufungidzira kuwanda kweprotein yakapihwa mu s.amples (zvinoenderana nemamwe sampzvishoma). Hunhu hwedhata kubva kune yega yega Ongorora kuteedzera kumhanya inonyanya kutsanangurwa neQC paramita yakasarudzika kune Olink tekinoroji. Nekudaro, akajairwa emhando yekudzora metrics anoshandiswa mune akajairwa NGS, senge Q-chibodzwa, haana kushomeka.

Bvisa uye urase katuriji mushure mekumhanya

YAMBIRO: Iyi seti yemareagents ine makemikari anogona kuva nengozi. Pfeka midziyo yekudzivirira yakakwana uye urase mareagents akashandiswa zvinoenderana nezviyero zvinoshanda. Kuti uwane rumwe ruzivo, tarisa kuIllumina NextSeq 1000 uye 2000 System Guide (gwaro #1000000109376).

1. Kana kumhanya kwapera, sarudza Eject Cartridge.
   • CHERECHEDZA: Iyo cartridge yakashandiswa inosanganisira kuyerera sero inogona kusiiwa munzvimbo kusvika pakumhanya kunotevera, asi kwete kupfuura mazuva matatu.
2. Bvisa cartridge kubva patireyi.
3. Rasa ma reagents zvinoenderana nemitemo inoshanda.
4. Sarudza Close Door. Iyo tray inoiswa zvakare.
5. Sarudza Home kuti udzokere kuHome screen.
   • CHERECHEDZA: Sezvo iyo cartridge ine nzira dzese dzekumhanyisa hurongwa, pamwe nedhiri rekuunganidza rakashandiswa reagents, hapana chikonzero chekugezesa chiridzwa mushure mekumhanya.

Nhoroondo yekudzokorora

Version	Date	Tsanangudzo
1.0	2021-12-01	New

www.olink.com

Zvekutsvaga Shandisa Chete. Kwete Yekushandisa muDiagnostic Procedus.

Ichi chigadzirwa chinosanganisira rezinesi rekusatengesa kushandiswa kweOlink zvigadzirwa. Vashandisi vezvokutengeserana vangada mamwe marezinesi. Ndokumbira ubate Olink Proteomics AB kuti uwane ruzivo. Iko hakuna waranti, inoratidzwa kana kurehwa, iyo inopfuurira kupfuura iyi tsananguro. Olink Proteomics AB haina mhosva yekukuvadzwa kwepfuma, kukuvara kwemunhu, kana kurasikirwa kwehupfumi kunokonzerwa nechigadzirwa ichi. Chiratidzo chinotevera ndeche Olink Proteomics AB: Olink®. Ichi chigadzirwa chakafukidzwa nematendi akati wandei uye patent application inowanikwa pa https://www.olink.com/patents/.

© Copyright 2021 Olink Proteomics AB. Ese matrademark echitatu-bato zvinhu zvevaridzi vawo.

Olink Proteomics, Dag Hammarskjölds väg 52B , SE-752 37 Uppsala, Sweden

1193, v1.0, 2021-12-01

Zvinyorwa / Zvishandiso

Olink NextSeq 2000 Ongorora Sequencing System [pdf] User Manual NextSeq 2000, Ongorora Sequencing System, NextSeq 2000 Ongorora Sequencing System

References

• User Manual