I-Olink NextSeq 2000 Hlola Uhlelo Lokulandelanisa

Inothi ledokhumenti

Imanuwali ye-Olink® Explore User, doc nr 1153, isiphelelwe yisikhathi, futhi ithathelwe indawo yimibhalo elandelayo:

• I-Olink® Hlola Ngaphezuluview Incwadi Yomsebenzisi, idokhumenti nr 1187
• I-Olink® Explore 384 User Manual, doc nr 1188
• I-Olink® Hlola i-4 x 384 Imanuwali yomsebenzisi, i-doc nr 1189
• I-Olink® Explore 1536 & Expansion User Manual, doc nr 1190
• I-Olink® Explore 3072 User Manual, doc nr 1191
• I-Olink® Hlola Ukulandelanisa usebenzisa i-NextSeq 550 User Manual, doc nr 1192
• I-Olink® Hlola Ukulandelanisa usebenzisa i-NextSeq 2000 User Manual, doc nr 1193
• I-Olink® Hlola Ukulandelanisa usebenzisa i-NovaSeq 6000 User Manual, doc nr 1194

Isingeniso

Ukusetshenziswa okuhlosiwe

I-Olink® Explore iyinkundla ye-multiplex immunoassay yokutholwa kwe-biomarker yamaprotheni omuntu. Umkhiqizo uhloselwe Ukusetshenziswa Kocwaningo Kuphela, futhi hhayi ukusetshenziswa ezinqubweni zokuxilonga. Umsebenzi waselabhorethri uzoqhutshwa kuphela abasebenzi baselabhorethri abaqeqeshiwe. Ukucubungula idatha kuzokwenziwa kuphela abasebenzi abaqeqeshiwe. Imiphumela ihloselwe ukusetshenziswa abacwaningi ngokuhlanganyela nokunye okutholwe emitholampilo noma elabhorethri.

Mayelana nale manuwali

Le Manuwali Yomsebenzisi inikeza imiyalelo edingekayo ukuze i-Olink® Explore Libraries ilandelelane ku-Illumina® NextSeq™ 2000. Imiyalo kufanele ilandelwe ngokuqinile nangokucacile. Noma yikuphi ukuchezuka kuzo zonke izinyathelo zaselabhorethri kungase kubangele idatha eyonakele. Ngaphambi kokuqala ukuhamba komsebenzi kulabhorethri, thintana ne-Olink® Explore Overview Incwadi Yomsebenzisi yesingeniso seplatifomu, okuhlanganisa ulwazi mayelana nama-reagents, okokusebenza kanye nemibhalo edingekayo, i-overview wokuhamba komsebenzi, kanye nemihlahlandlela yaselabhorethri. Ukuze uthole imiyalelo yokuthi uwasebenzise kanjani amakhithi e-Olink® Explore Reagent, bheka i-Olink® Explore User Manual esebenzayo. Ngokucubungula idatha nokuhlaziywa kwemiphumela yokulandelana kwe-Olink® Explore, bheka ku-Olink® MyData Cloud User Guide. Zonke izimpawu zokuthengisa namalungelo okushicilela aqukethwe kulolu hlelo lokusebenza kuyimpahla ye-Olink® Proteomics AB, ngaphandle uma kushiwo ngenye indlela.

Ukusekela kwezobuchwepheshe

Ukuze uthole ukwesekwa kobuchwepheshe, xhumana ne-Olink Proteomics ku: support@olink.com.

Iziyalezo zaselabhorethri

Lesi sahluko sinikeza imiyalelo yokuthi ungalandelanisa kanjani i-Olink Libraries ku-NextSeq™ 2000 usebenzisa i-NextSeq™ 1000/2000 P2 Reagents (100 Cycles) v3. Iphrothokholi esetshenziselwa ukulandelana iwukujwayela kokuhamba komsebenzi kwe-Illumina® NGS okujwayelekile kwe-Illumina® NextSeq™ 2000. Ngaphambi kokuqhubeka nokulandelana, qiniseka ukuthi ikhwalithi ye-Olink Library ehlanzekile iqinisekisiwe. Bheka i-Olink Explore User Manual esebenzayo ukuze uthole imiyalelo mayelana nokulawula ikhwalithi.

Hlela ukugijima kokulandelana

Ilabhulali eyodwa ye-Olink ingalandelaniswa ngeseli eligelezayo le-NextSeq™ 2000 P2 kanye nokugijima ngakunye. Inani lamaseli agelezayo e-P2 nokugijima okudingekayo ukuze kulandelelane amakhithi e-Olink Explore Reagent ahlukene kuchazwe kuThebula 1. Uma kudingeka ukugijima okungaphezu kokukodwa, phinda imiyalelo echazwe kule manuwali.

Ithebula 1. Ukulandelanisa ukuhlela kokugijima

Ikhithi ye-Olink® Hlola iReagent	Inombolo Yemitapo yolwazi ye-Olink	Inani lamaseli agelezayo kanye nokuqalisa
I-Olink® Explore 384 Reagent Kit	1	1
I-Olink® Explore 4 x 384 Reagent Kit	4	4
I-Olink® Explore 1536 Reagent Kit	4	4
Ikhithi ye-Olink® Explore Expansion Reagent	4	4
I-Olink® Explore 3072 Reagent Kit	8	8

Faka iresiphi yangokwezifiso ye-Olink®

Londoloza iresiphi yangokwezifiso ye-Olink xml-file Olink_NSQ2K_P2_V1 kufolda yensimbi efanele.

QAPHELA: Iresiphi yangokwezifiso ye-Olink izosebenza kuphela nekhithi ye-NextSeq™ 1000/2000 P2 Reagents (100 Cycles) v3 kanye ne-NextSeq™ 1000/2000 control software v1.2 noma v1.4.

Lungiselela ukulandelana kwama-reagents

Phakathi nalesi sinyathelo, i-cartridge ye-reagent equkethe ama-reagents e-clustering kanye nokulandelana iyancibilika futhi iseli eligelezayo liyalungiswa.

ISEXWAYISO: I-cartridge ye-reagent iqukethe amakhemikhali angaba yingozi. Gqoka izinto ezivikelayo ezanele futhi ulahle ama-reagents asetshenzisiwe ngokuhambisana namazinga asebenzayo. Ukuze uthole ulwazi olwengeziwe, bheka i-Illumina NextSeq 1000 kanye ne-2000 System Guide (idokhumenti #1000000109376).

Lungisa i-cartridge ye-reagent

Ukuncibilika kwe-cartridge engavuliwe kungenziwa ngokusebenzisa izindlela ezintathu ezihlukene: ekamelweni lokushisa, endaweni yokugeza yamanzi elawulwayo, noma esiqandisini.

Lungiselela ibhentshi

• I-1x NextSeq™ 1000/2000 P2 Reagent Cartridge (imijikelezo eyi-100)

Iziyalezo

1. Ncibilikisa i-cartridge ye-reagent njengoba kuchazwe kuThebula 2.

Ithebula 2. Izindlela zokuncibilikisa i-cartridge ye-reagent

Indlela yokupheka	Iziyalezo
Ngezinga lokushisa lekamelo	Ngaphandle kokuvula isikhwama se-foil esilisiliva, beka i-cartridge ye-reagent efriziwe ebhentshini bese uyincibilikisa ekamelweni lokushisa amahora angu-9. Ungadluli amahora angu-16. Qinisekisa ukuthi ilebula yesikhwama ibheke phezulu nokuthi umoya ungazungeza i-cartridge.
Ebhavini lamanzi	Ngaphandle kokuvula isikhwama se-foil esisiliva, faka i-cartridge ye-reagent eqandisiwe icwiliswe uhhafu kubhavu wamanzi olawulwayo ongu-25 °C futhi uyivumele inyibilike amahora angu-6. Ungadluli amahora angu-8. Qinisekisa ukuthi ilebula yesikhwama ibheke phezulu nokuthi i-cartridge ayiguquki ngesikhathi sokuncibilika. Yomisa i-cartridge kahle ngethawula lephepha.
Esiqandisini	Ngaphandle kokuvula isikhwama se-foil esiliva, beka i-cartridge ebhentshini bese uyincibilikisa ekamelweni lokushisa amahora angu-6. Qinisekisa ukuthi ilebula yesikhwama ibheke phezulu nokuthi umoya ungazungeza i-cartridge. Qedela ukuncibilikisa i-cartridge esiqandisini amahora angu-12. Ungadluli amahora angama-72. Ngaphambi kokugijima, susa i-cartridge engavulwanga encibilikisiwe esiqandisini futhi uyivumele ukuthi ifinyelele izinga lokushisa ekamelweni okungenani imizuzu engu-15, kodwa ingabi ngaphezu kwehora elingu-1.

QAPHELA: Ama-cartridge ancibilikisiwe awakwazi ukuphinda aqandiswe futhi kufanele agcinwe ku-4 °C, isikhathi esiphezulu esingamahora angama-72.

Lungiselela iseli eligelezayo

Lungiselela ibhentshi

• 1x NextSeq™ 1000/2000 P2 Flow Cell

Iziyalezo

1. Letha i-flow cell efrijini ekamelweni lokushisa imizuzu engu-10-15.

Lungiselela i-Olink® Library ukuze ilandelelane

Phakathi nalesi sinyathelo, i-Olink Library ehlanziwe nekhwalithi elawulwa yikhwalithi ihlanjululwa ekugxilweni kokugcina kokulayisha. Qaphela ukuthi ukuguqulwa kweLabhulali kwenziwa ngokuzenzakalelayo kuthuluzi.

Lungiselela ibhentshi

• I-Lib Tube, elungiselelwe ngokuya nge-Olink Explore User Manual esebenzayo
• 1x i-RSB ene-Tween 20
• Amanzi e-MilliQ
• 2x amashubhu e-Microcentrifuge (1.5 mL)
• I-pipette eyenziwe ngesandla (10, 100 kanye ne-1000 μL)
• Hlunga amathiphu we-pipette

Ngaphambi kokuthi uqale

• Ncibilikisa i-Lib Tube uma ifriziwe.
• Ncibilikisa i-RSB eqandisiwe nge-Tween 20 ekamelweni lokushisa imizuzu eyi-10. Gcina ku +4 °C uze usetshenziswe.
• Maka amashubhu amabili amasha angu-1.5 mL microcentrifuge kanje:
  • Maka ishubhu eyodwa ngokuthi “Dil” (kuMtapo wolwazi ohlanjululwe ongu-1:100)
  • Maka ishubhu elilodwa elithi “Seq” (ukuze lilungele ukulayisha Umtapowolwazi)

Iziyalezo

1. Engeza u-495 μL wamanzi we-MilliQ ku-Dil Tube.
2. Vortex the Lib Tube bese uyiphotha phansi kafushane.
3. Dlulisa ngesandla u-5 μL usuka ku-Lib Tube uye ku-Dil Tube.
4. Vortex the Dil Tube bese uyiphotha phansi kafushane.
5. Engeza u-20 μL we-RBS ne-Tween 20 ku-Seq Tube.
6. Dlulisa ngokwakho u-20 μL usuka ku-Dil Tube uye ku-Seq Tube.
7. Vortex the Seq Tube bese uyiphotha phansi kafushane.
8. Ngokushesha qhubekela ku-2.5 Load flow cell kanye ne-Olink® Library ku-cartridge ye-reagent.

QAPHELA: Gcina i-Lib Tube ku--20 °C uma kwenzeka kuphinda kuphindwe.

Layisha iseli eligelezayo kanye ne-Olink® Library ku-cartridge ye-reagent

Phakathi nalesi sinyathelo, iseli eligelezayo kanye ne-Olink Library ehlanjululwe kulayishwa kukhatriji ye-reagent encibilikisiwe.

Lungiselela ibhentshi

• I-1x encibilikisiwe i-NextSeq™ 1000/2000 P2 Reagent Cartridge (imijikelezo eyi-100), elungiswe esinyathelweni sangaphambilini
• I-1x NextSeq™ 1000/2000 P2 Flow Cell, elungiselelwe esinyathelweni sangaphambilini
• I-Seq Tube (esilungele ukulayisha i-Olink Library ehlanjululwe), elungiswe esinyathelweni sangaphambilini
• Ipayipi lesandla (100 μL)
• Ithiphu ye-Pipette (1 mL)

Lungisa i-cartridge

1. Susa i-cartridge esikhwameni sesiliva se-foil.
2. Guqula i-cartridge izikhathi eziyishumi ukuze uhlanganise kahle ama-reagents ancibilikisiwe ngaphakathi.

QAPHELA: Kuyinto evamile ukuzwa izingxenye zangaphakathi zishikiza.

Layisha i-flow cell ku-cartridge

1. Uma usulungele ukulayisha i-flow cell ku-cartridge, susa i-flow cell ephaketheni. Bamba iseli eligelezayo ngethebhu empunga, ilebula kuthebhu ibheke phezulu. Sebenzisa amagilavu ​​amasha angenayo impushana ukuze ugweme ukungcolisa indawo yengilazi yeseli eligelezayo.
2. Faka iseli lokugeleza ku-flow cell slot ngaphambili kwe-cartridge. Ukuchofoza okuzwakalayo kubonisa ukuthi iseli eligelezayo libekwe ngendlela efanele.
3. Susa ithebhu empunga ngokuyikhipha.

Layisha i-Olink® Library kukhathriji

1. Gcoba indawo yokugcina amanzi ngethiphu elihlanzekile elingu-1 mL.
2. Layisha u-20 μL we-Olink Library usuka ku-Seq Tube uye ngaphansi kwechibi lomtapo wezincwadi.

Yenza ukugijima kokulandelana kwe-Olink®

Phakathi nalesi sinyathelo, i-Buffer cartridge eneseli eligelezayo elilayishiwe kanye ne-Olink Library ilayishwa ku-NextSeq™ 2000, futhi ukuqalisa kokulandelana kuqalwa kusetshenziswa iresiphi ye-Olink.

Lungiselela ibhentshi

• I-1x NextSeq™ 1000/2000 P2 Reagent Cartridge (imijikelezo eyi-100) elayishwe nge-NextSeq™ 1000/2000 P2 Flow Cell kanye ne-Olink Library ehlanjululiwe, elungiswe esinyathelweni sangaphambilini.

Lungiselela i-Run Mode

1. Kumenyu yesofthiwe yokulawula, khetha Izilungiselelo.
2. Ngaphansi kwe-BaseSpace Sequence Hub Services kanye Nosekelo Olusebenzayo, khetha Ukusethwa Kwe-Local Run.
3. Khetha Ukusekela Okusebenzayo Kuphela njengezilungiselelo ezengeziwe. Lo msebenzi udinga uxhumano lwe-inthanethi.
4. Khetha Indawo Yokusingatha idatha yakho. Indawo yokusingatha kufanele ibe phakathi noma eduze nesifunda sakho.
5. Setha indawo Yefolda Yokukhiphayo ukuze uthole idatha esetshenziswayo yamanje. Khetha okuthi Khetha ukuze uzulazule bese ukhetha ifolda yokuphumayo.
6. Khetha ibhokisi lokuhlola le-Denature kanye ne-Dilute On Board ukuze uguqule ngokuzenzakalelayo futhi uhlambulule Ilabhulali egibele ithuluzi.
7. Khetha ibhokisi likaqhwishi le-Purge Reagent Cartridge ukuze uhlanze ngokuzenzakalelayo ama-reagents angasetshenzisiwe endaweni esetshenzisiwe esetshenzisiwe ye-cartridge.
8. Khetha ibhokisi lokuthikha elithi Hlola ngokuzenzakalela lezibuyekezo zesofthiwe ukuze uhlole ngokuzenzakalelayo izibuyekezo zesofthiwe (uyazikhethela). Lo msebenzi udinga uxhumano lwe-inthanethi.
9. Khetha Londoloza.

Setha amapharamitha wokugijima

QAPHELA: Lo myalelo usebenza enguqulweni 1.4 yesofthiwe yokulawula ye-NextSeq™ 1000/2000. Ezinye zezinyathelo ezichazwe ngezansi zingase zehluke uma usebenzisa inguqulo v1.2

1. Kumenyu yesofthiwe yokulawula, khetha Qala.
2. Khetha Mathupha Setha Ukuqalisa Okusha bese ucindezela u-Setha.
3. Ekhasini elithi Run Setup, setha amapharamitha alandelayo:
   • Esigabeni se-Run Name, faka i-ID yokuhlola eyingqayizivele.
   • Ohlwini lokudonsela phansi Lohlobo Lokufunda, khetha inketho yokufunda Okukodwa.
   • Faka inombolo yemijikelezo kanje:
     • Funda 1: 24
     • Inkomba 1: 0
     • Inkomba 2: 0
     • Funda 2: 0

QAPHELA: Kubalulekile ukuthi i-Read 1 isethwe ku-24, ngaphandle kwalokho konke ukugijima kuzohluleka. Ziba imilayezo yesixwayiso uma ufaka inombolo yemijikelezo.

• Ohlwini lokudonsela phansi lwe-Custom Primer Wells, khetha Cha.
• Kunkambu yeresiphi yangokwezifiso (uyazikhethela), khetha okuthi Khetha ukuzulazula bese ukhetha i-XML yeresiphi yangokwezifiso file Olink_NSQ2K_P2_V1. Khetha Vula.
• Ungangenisi i-Sample Sheet.
• Qiniseka ukuthi indawo Yefolda Yokukhiphayo ilungile. Uma kungenjalo, khetha okuthi Khetha ukuzulazula bese ukhetha indawo yefolda yokukhiphayo oyifunayo.
• Kunkambu ye-Denature and Dilute Onboard, khetha okuthi Kunikwe amandla ohlwini lokudonsela phansi.
• Khetha okuthi Lungisa.

Layisha i-cartridge elayishiwe

1. Khetha Layisha. I-visor yensimbi iyavuleka bese ithreyi liyakhishwa.
2. Beka i-cartridge elayishiwe ethreyini ilebula ibheke phezulu kanye neseli eligeleza ngaphakathi kwensimbi.
3. Khetha Vala.
4. Uma i-cartridge ilayishwe kahle, qinisekisa amapharamitha wokugijima bese ukhetha Ukulandelana. Ithuluzi lenza amasheke asetshenziswe ngaphambilini ensimbi kanye ne-fluidics.
   • QAPHELA: Ngesikhathi sokuhlolwa kwe-fluidics, kulindeleke ukuthi kuzwe imisindo eminingana ephumayo.
5. Qinisekisa ukuthi ukugijima kuqala ngemva kokuthi ukuhlola okuzenzakalelayo kokwenziwa ngaphambilini sekuqediwe (~imizuzu engu-15). Isikhathi sokusebenza esilandelanayo cishe yi-10h30 min.
   • QAPHELA: Nganoma yikuphi ukwehluleka kokuhlolwa kwangaphambilini, bheka imiyalelo yomkhiqizi. Qaphela ukuthi ungashayisani noma uphazamise i-NextSeq™ 2000 ngesikhathi sokulandelana kokuqalisa. Insimbi izwela ukudlidliza.
6. Hlanza indawo yokusebenza.

Gada Intuthuko Yokugijima

U-Olink usebenzisa i-NGS njengokufunda ukuze alinganise inani lokulandelana okwaziwayo ukuze alinganisele ukuhlangana kwephrotheni enikeziwe ku-s.amples (okuhlobene nezinye iziampkancane). Ikhwalithi yedatha evela ekugijimeni ngakunye kokulandelana kwe-Hlola inqunywa ikakhulukazi amapharamitha e-QC ahlukile kubuchwepheshe be-Olink. Ngakho-ke, amamethrikhi ajwayelekile okulawula ikhwalithi asetshenziswa ku-NGS evamile, njenge-Q-score, awabalulekile kangako.

Khipha futhi ulahle i-cartridge ngemva kokugijima

ISEXWAYISO: Leli sethi lama-reagents liqukethe amakhemikhali angaba yingozi. Gqoka izinto ezivikelayo ezanele futhi ulahle ama-reagents asetshenzisiwe ngokuhambisana namazinga asebenzayo. Ukuze uthole ulwazi olwengeziwe, bheka i-Illumina NextSeq 1000 kanye ne-2000 System Guide (idokhumenti #1000000109376).

1. Uma ukugijima sekuqedile, khetha Khipha i-Cartridge.
   • QAPHELA: I-cartridge esetshenzisiwe ehlanganisa i-flow cell ingashiywa endaweni kuze kube yilapho igijima elandelayo, kodwa kungabi ngaphezu kwezinsuku ezi-3.
2. Susa i-cartridge kuthreyi.
3. Lahla ama-reagents ngokuhambisana namazinga asebenzayo.
4. Khetha okuthi Vala Umnyango. Ithreyi lilayishwa kabusha.
5. Khetha Ikhaya ukuze ubuyele esikrinini sasekhaya.
   • QAPHELA: Njengoba i-cartridge iqukethe zonke izindlela zokuqhuba uhlelo, kanye nedamu lokuqoqa ama-reagents asetshenzisiwe, asikho isidingo sokugeza insimbi ngemva kokugijima.

Umlando wokubuyekeza

Inguqulo	Usuku	Incazelo
1.0	2021-12-01	Okusha

www.olink.com

Okokusetshenziswa Kocwaningo Kuphela. Ayisetshenziselwa Izinqubo Zokuxilonga.

Lo mkhiqizo uhlanganisa ilayisense yokusebenzisa okungekona okokuthengisa kwemikhiqizo ye-Olink. Abasebenzisi bezentengiso bangadinga amalayisense engeziwe. Sicela uthinte i-Olink Proteomics AB ukuze uthole imininingwane. Azikho iziqinisekiso, eziveziwe noma ezishiwo, ezidlula le ncazelo. I-Olink Proteomics AB ayinasibopho sokulimala kwempahla, ukulimala komuntu siqu, noma ukulahlekelwa kwezomnotho okubangelwe yilo mkhiqizo. Uphawu lokuhweba olulandelayo luphethwe yi-Olink Proteomics AB: Olink®. Lo mkhiqizo umbozwe ngamalungelo obunikazi amaningana kanye nezicelo zelungelo lobunikazi ezitholakala lapha https://www.olink.com/patents/.

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

1193, v1.0, 2021-12-01

Amadokhumenti / Izinsiza

I-Olink NextSeq 2000 Hlola Uhlelo Lokulandelanisa [pdf] Imaniwali yosebenzisayo I-NextSeq 2000, Hlola Uhlelo Lokulandelanisa, i-NextSeq 2000 Hlola Uhlelo Lokulandelanisa

Izithenjwa

• Imaniwali yosebenzisayo