Olink NextSeq 2000 Phonononga iSistim yolandelelwano

Inqaku loxwebhu

I-Olink® Explore User manual, doc nr 1153, ayisebenzi, kwaye ithatyathelwe indawo ngala maxwebhu alandelayo:

• Olink® Phonononga ngaphayaview Incwadi yoMsebenzisi, idoc nr 1187
• I-Olink® Phonononga i-384 yeNcwadi yoMsebenzisi, i-doc nr 1188
• Olink® Phonononga 4 x 384 Incwadi yoMsebenzisi, idoc nr 1189
• I-Olink® Phonononga i-1536 kunye neNcwadi yoMsebenzisi yoKwandiswa, idoc nr 1190
• I-Olink® Phonononga i-3072 yeNcwadi yoMsebenzisi, i-doc nr 1191
• I-Olink® Phonononga ulandelelwano usebenzisa i-NextSeq 550 Incwadi Yomsebenzisi, idoc nr 1192
• I-Olink® Phonononga ulandelelwano usebenzisa i-NextSeq 2000 Incwadi Yomsebenzisi, idoc nr 1193
• I-Olink® Phonononga ulandelelwano usebenzisa iNovaSeq 6000 Manual yomsebenzisi, doc nr 1194

Intshayelelo

Ukusetyenziswa okucetywayo

I-Olink® Phonononga liqonga le-multiplex immunoassay lokufumanisa iprotein biomarker yomntu. Imveliso yenzelwe ukuSebenza koPhando kuphela, kwaye kungekhona ukusetyenziswa kwiinkqubo zokuxilonga. Umsebenzi waselabhoratri uya kuqhutywa kuphela ngabasebenzi baselabhoratri abaqeqeshiweyo. Ukusetyenzwa kwedatha kuya kwenziwa kuphela ngabasebenzi abaqeqeshiweyo. Iziphumo zenzelwe ukuba zisetyenziswe ngabaphandi ngokubambisana nezinye iziphumo zeklinikhi okanye elebhu.

Malunga nale ncwadana

Le Ncwadi yoMsebenzisi ibonelela ngemiyalelo efunekayo ukulandelelanisa i-Olink® Explore Libraries kwi-Illumina® NextSeq™ 2000. Imiyalelo mayilandelwe ngokungqongqo nangokucacileyo. Nakuphi na ukutenxa kuwo onke amanyathelo elabhoratri kunokubangela ukonakaliswa kwedatha. Phambi kokuba uqalise ukuhamba kwelabhoratri, jongana ne-Olink® Explore Overview Incwadi yoMsebenzisi yentshayelelo eqongeni, kubandakanya ulwazi malunga neerejenti, izixhobo kunye namaxwebhu afunekayo, ngaphezuluview wokuhamba komsebenzi, kunye nezikhokelo zaselabhoratri. Ukufumana imiyalelo malunga nendlela yokusebenzisa i-Olink® Explore Reagent Kits, bhekisa kwi-Olink® Explore User Manual esebenzayo. Ukulungiswa kwedatha kunye nohlalutyo lwe-Olink® Hlola iziphumo zokulandelelana, bhekisa kwi-Olink® MyData Cloud User Guide. Zonke iimpawu zokuthengisa kunye namalungelo okushicilela aqulethwe kwesi sixhobo yipropati ye-Olink® Proteomics AB, ngaphandle kokuba kuchazwe ngenye indlela.

Uxhaso lobuchwepheshe

Ngenkxaso yobugcisa, qhagamshelana ne-Olink Proteomics apha: support@olink.com.

Imiyalelo yeLebhu

Esi sahluko sibonelela ngemiyalelo ngendlela yolandelelwano lweeLayibrari ze-Olink kwi-NextSeq™ 2000 usebenzisa i-NextSeq™ 1000/2000 P2 Reagents (i-100 Cycles) v3. Iprotocol esetyenziselwa ukulandelelanisa i-adaptation ye-Illumina® standard NGS workflow for Illumina® NextSeq™ 2000. Ngaphambi kokuba uqhubeke nokulandelelana, qiniseka ukuba umgangatho we-Olink Library ococekileyo uqinisekisiwe. Jonga kwi-Olink esebenzayo Phonononga Incwadi Yomsebenzisi ngemiyalelo malunga nolawulo lomgangatho.

Cwangcisa ukubaleka kolandelelwano

Ithala leencwadi le-Olink elinye linolandelelwano ngokwe-NextSeq™ 2000 P2 flow cell kunye nokubaleka ngakunye. Inani leeseli ezijikelezayo ze-P2 kunye nokuqhuba okufunekayo ukulandelelanisa i-Olink Explore Reagent Kits ichazwe kwiThebhile 1. Ukuba kufuneka ukubaleka okungaphezulu kwesinye, phinda imiyalelo echazwe kule ncwadana.

Uluhlu 1. Ukulandelelanisa isicwangciso sokubaleka

Olink® Phonononga iReagent Kit	Inani leelayibrari ze-Olink	Inani lokuqukuqela (ii)seli kunye ne(s)
I-Olink® Phonononga i-384 Reagent Kit	1	1
I-Olink® Phonononga i-4 x 384 yeReagent Kit	4	4
I-Olink® Phonononga i-1536 Reagent Kit	4	4
Olink® Phonononga Ukwandiswa kweReagent Kit	4	4
I-Olink® Phonononga i-3072 Reagent Kit	8	8

Faka iresiphi yesiqhelo ye-Olink®

Gcina iresiphi yesiko le-Olink xml-file Olink_NSQ2K_P2_V1 kwifolda yesixhobo esifanelekileyo.

PHAWULA: I-recipe yesiko le-Olink iya kusebenza kuphela nge-NextSeq™ 1000/2000 P2 Reagents (i-100 Cycles) v3 kit kunye ne-NextSeq™ 1000/2000 yokulawula isoftware v1.2 okanye v1.4.

Lungiselela ulandelelwano reagents

Ngethuba le nyathelo, i-cartridge ye-reagent equkethe i-clustering kunye ne-reagents yokulandelelanisa iyanyibilika kwaye iseli yokuhamba iyalungiswa.

ISILUMKISO: Ikhatriji yereagent iqulethe iikhemikhali ezinokuba yingozi. Nxiba izixhobo ezifanelekileyo zokukhusela kwaye ulahle ii-reagents ezisetyenzisiweyo ngokuhambelana nemigangatho esebenzayo. Ngolwazi oluthe kratya, bhekisa kwi-Illumina NextSeq 1000 kunye ne-2000 System Guide (uxwebhu #1000000109376).

Lungisa ikhatriji yereagent

Ukunyibilika kwe-cartridge engavulwanga kunokwenziwa ngeendlela ezintathu ezahlukeneyo: kwiqondo lokushisa, kwindawo yokuhlambela yamanzi elawulwayo, okanye kwifriji.

Lungisa ibhentshi

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

Imiyalelo

1. Tyibilikisa ikhatriji ye-reagent njengoko kuchaziwe kwiTheyibhile yesi-2.

Itheyibhile 2. Iindlela zokunyibilikisa ikhatriji yeReagent

Indlela yokunyibilika	Imiyalelo
Kwiqondo lobushushu begumbi	Ngaphandle kokuvula ibhegi yefoyile yesilivere, beka i-cartridge ye-reagent ekhenkcezileyo kwibhentshi kwaye uyinyibilikise kwindawo yobushushu begumbi iiyure ezili-9. Ungagqithi kwiiyure ezili-16. Qinisekisa ukuba ileyibhile yengxowa ijonge phezulu kwaye umoya unokujikeleza kwikhatriji.
Kwibhafu yamanzi	Ngaphandle kokuvula ibhegi yefoyile yesilivere, beka ikhatriji engumkhenkce entywiliselwe kwindawo elawulwayo yokuhlamba amanzi eyi-25 °C kwaye uyiyeke inyibilike iiyure ezi-6. Ungagqithi kwiiyure ezisi-8. Qinisekisa ukuba ileyibhile yengxowa ijonge phezulu kwaye ikhatriji ayijiki ngexesha lokunyibilika. Yomisa i-cartridge ngokucokisekileyo ngetawuli yephepha.
Kwifriji	Ngaphandle kokuvula isikhwama se-foil yesilivere, faka i-cartridge kwibhentshi kwaye uyinyibilikise kwindawo yokushisa kweeyure ezingama-6. Qinisekisa ukuba ileyibhile yengxowa ijonge phezulu kwaye umoya unokujikeleza kwikhatriji. Ukugqiba ukunyibilikisa i-cartridge kwifriji yeeyure ze-12. Ungagqithi kwiiyure ezingama-72. Ngaphambi kokuba ubaleke, susa i-cartridge engavulwanga edibeneyo kwifriji kwaye uyivumele ukuba ifikelele kwiqondo lokushisa kwegumbi ubuncinane ubuncinane bemizuzu eyi-15, kodwa ingabi ngaphezu kweyure eli-1.

PHAWULA: Iikhatriji ezinyibilikisiweyo azikwazi ukuphinda zifakwe emkhenkceni kwaye kufuneka zigcinwe kwi-4 °C, ixesha elide leeyure ezingama-72.

Lungisa i-flow cell

Lungisa ibhentshi

• 1x NextSeq™ 1000/2000 iSeli eQuquzelayo yeP2

Imiyalelo

1. Yizisa i-flow cell efrijini kwindawo yokushisa kwe-10-15 imizuzu.

Lungiselela iThala leencwadi le-Olink® ukulungiselela ulandelelwano

Ngeli nyathelo, iThala leencwadi le-Olink elihlanjululweyo kunye nomgangatho olawulwayo lihlanjululwe kugxininiso lokugqibela lokulayisha. Qaphela ukuba ukuchaswa kweThala leencwadi kwenziwa ngokuzenzekelayo kwisixhobo.

Lungisa ibhentshi

• I-Lib Tube, elungiselelwe ngokuhambelana ne-Olink yokuhlola i-Manual yoMsebenzisi
• 1x i-RSB ene-Tween 20
• MilliQ amanzi
• Iityhubhu ezi-2x zeMicrocentrifuge (1.5 mL)
• Ipayipi yesandla (10, 100 kunye ne-1000 μL)
• Iingcebiso ze-pipette zokucoca

Ngaphambi kokuba uqale

• Nyibilikisa iLib Tube ukuba ingumkhenkce.
• Thaw i-RSB ekhenkcezisiweyo kunye ne-Tween 20 kwiqondo lobushushu begumbi imizuzu eyi-10. Gcina ku +4 °C de usetyenziswe.
• Phawula iityhubhu ezimbini ze-1.5 mL ze-microcentrifuge ngolu hlobo lulandelayo:
  • Phawula ityhubhu enye “Dil” (yeThala leencwadi elidilulweyo le-1:100)
  • Phawula ityhubhu enye “Seq” (ukulungiselela ukulayisha iThala leencwadi)

Imiyalelo

1. Yongeza i-495 μL yamanzi e-MilliQ kwi-Dil Tube.
2. Vortex iLib Tube kwaye uyijikeleze ngokufutshane.
3. Utshintshe ngesandla i-5 μL ukusuka kwiLib Tube ukuya kwiThubhu yeDil.
4. Vortex i-Dil Tube kwaye uyijikeleze ngokufutshane.
5. Yongeza i-20 μL ye-RBS kunye ne-Tween 20 kwi-Seq Tube.
6. Utshintshe ngesandla i-20 μL ukusuka kwiThubhu yeDil ukuya kwiThubhu yeSeq.
7. Vortex i-Seq Tube kwaye uyijikeleze ngokufutshane.
8. Ngokukhawuleza qhubeleka kwi-2.5 Load flow cell kunye ne-Olink® Library kwikhatriji ye-reagent.

PHAWULA: Gcina iLib Tube (s) kwi -20 °C xa kunokwenzeka u(zi)phinda-phinda.

Layisha iseli kunye nethala leencwadi le-Olink® kwikhatriji ye-reagent

Ngeli nyathelo, iseli yokuhamba kunye neLayibrari ye-Olink ehlanjululweyo ilayishwa kwi-cartridge ye-reagent encibilikisiweyo.

Lungisa ibhentshi

• I-1x enyibilikisiwe i-NextSeq™ 1000/2000 ye-P2 Reagent Cartridge (imijikelo eli-100), elungiswe kwinyathelo elidlulileyo
• I-1x NextSeq™ 1000/2000 P2 Flow Cell, elungiselelwe kwinqanaba elidlulileyo
• I-Seq Tube (elungele ukulayisha iThala leencwadi le-Olink elihlanjululweyo), elungiselelwe kwinqanaba langaphambili
• Ipayipi yesandla (100 μL)
• Ingcebiso yePipette (1 mL)

Lungisa i-cartridge

1. Susa i-cartridge kwisikhwama se-foil yesilivere.
2. Guqula ikhatriji kalishumi ukuze udibanise ngokucokisekileyo ii-reagents ezinyibilikisiweyo ngaphakathi.

PHAWULA: Yinto eqhelekileyo ukuva amalungu angaphakathi etswina.

Layisha iseli yokuhamba kwikhatriji

1. Xa ulungele ukulayisha i-flow cell kwi-cartridge, susa i-flow cell kwi-package. Bamba iseli engwevu, neleyibhile ekwithebhu ijonge phezulu. Sebenzisa iiglavu zasimahla zomgubo ukunqanda ukungcolisa umphezulu weglasi yeseli yokuhamba.
2. Faka i-flow cell kwi-flow cell slot ngaphambili kwekhatriji. Ukucofa okuvakalayo kubonisa ukuba i-flow cell ibekwe ngokuchanekileyo.
3. Susa ithebhu engwevu ngokuyikhupha.

Layisha iLayibrari ye-Olink® kwikhatriji

1. Gcoba ithala lethala leencwadi ngencam ecocekileyo ye-1 mL yepipette.
2. Layisha i-20 μL yeThala leeNcwadi le-Olink ukusuka kwiThubhu ye-Seq ukuya ezantsi kwindawo yokugcina amanzi.

Yenza i-Olink® yokulandelelanisa ukubaleka

Ngeli nyathelo, i-Buffer cartridge ene-flow flow cell kunye ne-Olink Library ilayishwe kwi-NextSeq™ 2000, kwaye ulandelelwano lokuqhuba luqaliswa kusetyenziswa iresiphi ye-Olink.

Lungisa ibhentshi

• I-1x i-NextSeq™ 1000/2000 ye-P2 Reagent Cartridge (imijikelo eyi-100) elayishwe nge-NextSeq™ 1000/2000 P2 Flow Cell kunye neThala leeNcwadi le-Olink elihlanjululweyo, elilungiselelwe kwinyathelo langaphambili.

Qwalasela iNdlela yokuSebenza

1. Kwimenyu yolawulo lwesoftwe, khetha Useto.
2. Ngaphantsi kwe-BaseSpace Sequence Hub Services & Proactive Support, khetha i-Local Run Setup.
3. Khetha iNkxaso eSebenzayo Kuphela njengeseto ezongezelelweyo. Lo msebenzi ufuna uqhagamshelo lwe-intanethi.
4. Khetha indawo yokusingatha idatha yakho. Indawo yokusingatha kufuneka ibe ngaphakathi okanye ikufutshane nommandla wakho.
5. Cwangcisa indawo yesiqulathi seefayili sesiphumo sangoku sedatha ekrwada. Khetha Khetha ukukhangela kwaye ukhethe ifolda yemveliso.
6. Khetha i-Denature kunye ne-Dilute kwiBhodi yokutshekisha ukuze ujike ngokuzenzekelayo kwaye unciphise iThala leencwadi kwisixhobo.
7. Khetha ibhokisi yokukhangela yeKhatriji yeReagent yokucoca ukuze uzicocise ngokuzenzekelayo ii-reagents ezingasetyenziswanga kwikhompatimenti ye-reagents echithiweyo yekhatriji.
8. Khetha i-AutoCheck yohlaziyo lwesoftwe ibhokisi yokukhangela ngokuzenzekelayo uhlaziyo lwesoftware (uyazikhethela). Lo msebenzi ufuna uqhagamshelo lwe-intanethi.
9. Khetha Gcina.

Cwangcisa iiparamitha zokuqhuba

PHAWULA: Lo myalelo usebenza kuguqulelo 1.4 lwesoftware yolawulo ye-NextSeq™ 1000/2000. Amanye ala manyathelo achazwe ngezantsi anokwahluka xa usebenzisa inguqulelo v1.2

1. Kwimenyu yesoftwe yolawulo, khetha Qala.
2. Khetha Ngokuzenzela Ukuseta uMsebenzi oMtsha kwaye ucinezele ukuSeta.
3. KwiPhepha lokuSebenzisa ukuSeta, seta iiparamitha zokuqhuba ngolu hlobo lulandelayo:
   • Kwindawo ethi Qhuba Igama, ngenisa i-ID yovavanyo olulodwa.
   • Kuludwe lokuthotywa kohlobo lokuFunda, khetha ukhetho lokuFunda okuNye.
   • Faka inani lemijikelo ngolu hlobo lulandelayo:
     • Funda 1: 24
     • Isalathiso 1: 0
     • Isalathiso 2: 0
     • Funda 2: 0

PHAWULA: Kubalulekile ukuba i-Read 1 imiselwe ku-24, kungenjalo yonke into iya kusilela. Ungayihoyi imiyalezo yesilumkiso xa ufaka inani lemijikelo.

• Kuluhlu lokuhla lwe-Custom Primer Wells, khetha Hayi.
• Kwindawo yeRecipe yesiko (ukhetho), khetha Khetha ukuhamba kwaye ukhethe iresiphi ye-XML yesiko file Ikhonkco_NSQ2K_P2_V1. Khetha Vula.
• Musa ukuthatha ngaphandle i-Sample Sheet.
• Qinisekisa ukuba indawo yesiqulathi seemveliso ichanekile. Kungenjalo, khetha Khetha ukukhangela kwaye ukhethe indawo efunekayo yesiqulathi seefayili.
• Kwi-Denature kunye ne-Dilute Onboard field, khetha Inikwe amandla kuluhlu oluhlayo.
• Khetha Lungisa.

Layisha ikhatriji elayishiweyo

1. Khetha Layisha. I-visor yesixhobo iyavula kwaye itreyi iyakhutshwa.
2. Beka i-cartridge elayishiwe kwi-tray kunye neleyibhile ejonge phezulu kunye neseli yokuhamba ngaphakathi kwesixhobo.
3. Khetha Vala.
4. Emva kokuba i-cartridge ilayishwe ngokufanelekileyo, qinisekisa iiparameters zokuqhuba kwaye ukhethe Ukulandelelana. Isixhobo senza iitshekhi zangaphambili zesixhobo kunye ne-fluidics.
   • PHAWULA: Ngexesha lokujonga i-fluidics, kulindeleke ukuba kuve izandi ezininzi eziphumayo.
5. Qinisekisa ukuba i-run iqala emva kokuba ukuhlolwa kwangaphambili okuzenzekelayo kugqityiwe (~ imizuzu eyi-15). Ixesha lokuqhuba ngokulandelelana limalunga ne-10h30 imizuzu.
   • PHAWULA: Nakuphi na ukusilela ekutshekishweni kwangaphambili, bhekisa kwimiyalelo yomenzi. Lumka ungangqubani okanye uphazamise i-NextSeq™ 2000 ngexesha lolandelelwano lokuqhuba. Isixhobo sinobuntununtunu kwintshukumo.
6. Coca indawo yokusebenza.

Beka iliso kwinkqubela phambili yokuqhuba

U-Olink usebenzisa i-NGS njenge-readout ukulinganisa inani lolandelelwano olwaziwayo ukuze uqikelele ukuxinwa kweprotheni enikeziweyo kwi-s.amples (ngokunxulumene nezinye iziqenduampkancinci). Umgangatho wedatha ukusuka nganye Phonononga ulandelelwano oluqhutywayo lumiselwa ikakhulu yiparamitha ye-QC ekhethekileyo kwitekhnoloji ye-Olink. Ke ngoko, iimetriki zolawulo lomgangatho osemgangathweni osetyenziswa kwi-NGS eqhelekileyo, njenge-Q-score, ayibalulekanga kangako.

Khipha kwaye ulahle i-cartridge emva kokubaleka

ISILUMKISO: Le seti yee-reagents iqulethe iikhemikhali ezinokuba yingozi. Nxiba izixhobo ezifanelekileyo zokukhusela kwaye ulahle ii-reagents ezisetyenzisiweyo ngokuhambelana nemigangatho esebenzayo. Ngolwazi oluthe kratya, bhekisa kwi-Illumina NextSeq 1000 kunye ne-2000 System Guide (uxwebhu #1000000109376).

1. Xa ukubaleka kugqityiwe, khetha Khupha ikhatriji.
   • PHAWULA: Ikhatriji esetyenzisiweyo kuquka i-flow cell inokushiywa kwindawo kude kube kuqhutywe okulandelayo, kodwa akukho ngaphezu kweentsuku ezi-3.
2. Susa ikhatriji kwitreyi.
3. Ukulahla i-reagents ngokuhambelana nemigangatho esebenzayo.
4. Khetha Vala ucango. Itreyi iyalayishwa kwakhona.
5. Khetha eKhaya ukubuyela kwiscreen sasekhaya.
   • PHAWULA: Ekubeni i-cartridge iqulethe zonke iindlela zokuqhuba inkqubo, kunye ne-reservoir yokuqokelela i-reagents esetyenzisiweyo, akukho mfuneko yokuhlamba isixhobo emva kokubaleka.

Imbali yohlaziyo

Inguqulelo	Umhla	Inkcazo
1.0	2021-12-01	Entsha

www.olink.com

Ukusetyenziswa koPhando kuphela. Ayinakusetyenziswa kwiiNkqubo zokuDinga.

Le mveliso ibandakanya ilayisenisi yokusetyenziswa okungeyontengiso yeemveliso ze-Olink. Abasebenzisi borhwebo banokufuna iilayisensi ezongezelelweyo. Nceda uqhagamshelane ne-Olink Proteomics AB ngeenkcukacha. Akukho ziqinisekiso, ezichaziweyo okanye ezichaziweyo, ezidlulela ngaphaya kwale nkcazo. I-Olink Proteomics AB ayinaxanduva lomonakalo wepropati, ukulimala komntu, okanye ukulahlekelwa kwezoqoqosho okubangelwa yile mveliso. Olu phawu lwentengiso lulandelayo luphethwe yi-Olink Proteomics AB: Olink®. Le mveliso igqunywa ngamalungelo awodwa omenzi wechiza kunye nezicelo zepatent ezikhoyo https://www.olink.com/patents/.

© Copyright 2021 Olink Proteomics AB. Zonke iimpawu zorhwebo zomntu wesithathu ziyipropati yabanini bazo.

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

1193, v1.0, 2021-12-01

Amaxwebhu / Izibonelelo

Olink NextSeq 2000 Phonononga iSistim yolandelelwano [pdf] Incwadi yokusebenzisa I-NextSeq 2000, Phonononga iNkqubo yolandelelwano, i-NextSeq 2000 Phonononga inkqubo yolandelelwano

Iimbekiselo

• Incwadi yokusebenzisa