Olink NextSeq 2000 Explore Sequencing System

Mongolo oa litokomane

Buka ea Olink® Explore User manual, doc nr 1153, ha e sa sebetsa, 'me e nkeloe sebaka ke litokomane tse latelang:

• Olink® Explore Overview Buka ea mosebelisi, doc nr 1187
• Olink® Explore 384 User Manual, doc nr 1188
• Olink® Explore 4 x 384 User Manual, doc nr 1189
• Olink® Explore 1536 & Expansion User Manual, doc nr 1190
• Olink® Explore 3072 User Manual, doc nr 1191
• Olink® Explore Sequencing sebelisa NextSeq 550 User Manual, doc nr 1192
• Olink® Explore Sequencing sebelisa NextSeq 2000 User Manual, doc nr 1193
• Olink® Explore Sequencing sebelisa NovaSeq 6000 User Manual, doc nr 1194

Selelekela

Tšebeliso e reriloeng

Olink® Explore ke sethala sa multiplex immunoassay bakeng sa ho sibolloa ha protheine ea motho. Sehlahisoa se reretsoe ho Sebelisa Lipatlisiso Feela, eseng ho sebelisoa mekhoeng ea ho hlahloba. Mosebetsi oa laboratori o tla etsoa feela ke basebetsi ba koetlisitsoeng ba laboratori. Ts'ebetso ea data e tla etsoa feela ke basebetsi ba koetlisitsoeng. Liphetho li reretsoe ho sebelisoa ke bafuputsi hammoho le liphuputso tse ling tsa kliniki kapa tsa laboratori.

Mabapi le bukana ena

Bukana ena ea Mosebelisi e fana ka litaelo tse hlokahalang ho hlophisa Olink® Explore Libraries ho Illumina® NextSeq™ 2000. Litaelo li tlameha ho lateloa ka tieo le ka mokhoa o hlakileng. Liphapang life kapa life mehatong eohle ea laboratori li ka baka ho senyeha ha data. Pele o qala tšebetso ea laboratori, ikopanye le Olink® Explore Overview Buka ea mosebelisi bakeng sa kenyelletso ea sethala, ho kenyelletsa le tlhahisoleseling mabapi le li-reagents, lisebelisoa le litokomane tse hlokahalang, ho fetaview ea tsamaiso ea mosebetsi, hammoho le tataiso ea laboratori. Bakeng sa litaelo tsa ho sebelisa Olink® Explore Reagent Kits, sheba Buka e sebetsang ea Olink® Explore User Manual. Bakeng sa ts'ebetso ea data le tlhahlobo ea liphetho tsa tatellano ea Olink® Explore, sheba ho Olink® MyData Cloud User Guide. Matšoao ohle a khoebo le litokelo tsa molao tse teng ka har'a sesebelisoa sena ke thepa ea Olink® Proteomics AB, ntle le haeba ho boletsoe ka tsela e 'ngoe.

Tšehetso ea tekheniki

Bakeng sa tšehetso ea tekheniki, ikopanye le Olink Proteomics ho: support@olink.com.

Litaelo tsa laboratori

Khaolo ena e fana ka litaelo tsa hore na u ka tatellana joang Lilaebrari tsa Olink ho NextSeq™ 2000 u sebelisa NextSeq™ 1000/2000 P2 Reagents (100 Cycles) v3. Protocol e sebelisetsoang ho latellana ke ho ikamahanya le Illumina® standard NGS workflow bakeng sa Illumina® NextSeq™ 2000. Pele u tsoela pele ka tatellano, etsa bonnete ba hore boleng ba Olink Library bo hloekisitsoeng bo tiisitsoe. Sheba bukana e sebetsang ea Olink Explore User Manual bakeng sa litaelo mabapi le taolo ea boleng.

Etsa moralo oa ho latellana

Laeborari e le 'ngoe ea Olink e ka hlophisoa ka seleng ea phallo ea NextSeq™ 2000 P2 le ho matha. Palo ea lisele tse phallang tsa P2 le li-run tse hlokahalang ho tatellano ea li-Olink Explore Reagent Kits li hlalositsoe ho Lethathamo la 1. Haeba ho hlokahala ho matha ho feta e le 'ngoe, pheta litaelo tse hlalositsoeng bukeng ena.

Lethathamo la 1. Ho hlophisa moralo oa ho matha

Olink® Explore Reagent Kit	Palo ea Lilaebrari tsa Olink	Nomoro ea (li)cell cell le ho matha
Olink® Explore 384 Reagent Kit	1	1
Olink® Explore 4 x 384 Reagent Kit	4	4
Olink® Explore 1536 Reagent Kit	4	4
Olink® Explore Expansion Reagent Kit	4	4
Olink® Explore 3072 Reagent Kit	8	8

Kenya risepe ea tloaelo ea Olink®

Boloka risepe ea tloaelo ea Olink xml-file Olink_NSQ2K_P2_V1 sephuthelong se nepahetseng sa sesebelisoa.

HLOKOMELA: Recipe ea tloaelo ea Olink e tla sebetsa feela le NextSeq™ 1000/2000 P2 Reagents (100 Cycles) v3 kit le NextSeq™ 1000/2000 control software v1.2 kapa v1.4.

Lokisetsa tatellano ea li-reagents

Nakong ea mohato ona, cartridge ea reagent e nang le li-clustering le li-reagents tse latellanang lia qhibiliha 'me sele ea phallo e lokisoa.

TLHOKOMELISO: Cartridge ea reagent e na le lik'hemik'hale tse ka bang kotsi. Apara lisebelisoa tse lekaneng tsa ts'ireletso 'me u lahle li-reagents tse sebelisitsoeng ho latela litekanyetso tse sebetsang. Bakeng sa tlhaiso-leseling e batsi, sheba ho Illumina NextSeq 1000 le 2000 System Guide (tokomane #1000000109376).

Lokisetsa khatriji ea reagent

Ho qhibiliha cartridge e sa buloang ho ka etsoa ka mekhoa e meraro e fapaneng: mocheso oa kamore, ka bateng ea metsi e laoloang, kapa ka sehatsetsing.

Lokisetsa benche

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

Litaelo

1. Qhibiliha khatriji ea reagent joalo ka ha ho hlalositsoe ho Lethathamo la 2.

Letlapa la 2. Mekhoa ea ho qhibiliha ea cartridge ea Reagent

Mokhoa oa ho qhibiliha	Litaelo
Ka mocheso oa kamore	Ntle le ho bula mokotla oa silevera, beha cartridge e nang le serame holim'a benche ebe u e qhibilihisa ka mocheso oa kamore bakeng sa lihora tse 9. Se ke oa feta lihora tse 16. Etsa bonnete ba hore leibole ea mokotla e shebane holimo le hore moea o ka potoloha khatriji.
Ka bate ea metsi	Ntle le ho bula mokotla oa silevera, beha khatriji e hatselitsoeng e qoelisoa ka har'a bate ea metsi e laoloang ea 25 °C 'me u e tlohele e qhibilihe lihora tse 6. Se ke oa feta lihora tse 8. Etsa bonnete ba hore leibole ea mokotla e shebile holimo le hore khatriji ha e khelohe nakong ea ho qhibiliha. Omisa cartridge hantle ka thaole ea pampiri.
Ka sehatsetsing	Ntle le ho bula mokotla oa silevera, beha cartridge bencheng ebe u e qhibilihisa mocheso oa kamore bakeng sa lihora tse 6. Etsa bonnete ba hore leibole ea mokotla e shebane holimo le hore moea o ka potoloha khatriji. Qetella ho qhibiliha cartridge ka sehatsetsing ka lihora tse 12. Se ke oa feta lihora tse 72. Pele o matha, tlosa cartridge e sa buloang ka sehatsetsing 'me u e lumelle hore e fihle mocheso oa kamore ka bonyane metsotso e 15, empa e se ke ea feta 1 hora.

HLOKOMELA: Likhatriji tse qhibilihisitsoeng ha li khone ho hoamisoa hape 'me li tlameha ho bolokoa ho 4 °C, nako e telele ea lihora tse 72.

Lokisetsa sele ea phallo

Lokisetsa benche

• 1x NextSeq™ 1000/2000 P2 Flow Cell

Litaelo

1. Tlisa sele ea phallo ea sehatsetsing mocheso oa kamore bakeng sa metsotso e 10-15.

Lokisetsa Olink® Library bakeng sa tatellano

Nakong ea mohato ona, Laebrari ea Olink e hloekisitsoeng le e laoloang ea boleng e hlapolloa ho fihlela mahloriso a ho qetela. Hlokomela hore denaturation ea Library e etsoa ka bo eona ka har'a sesebelisoa.

Lokisetsa benche

• Lib Tube, e lokiselitsoeng ho latela Bukana e sebetsang ea Olink Explore User
• 1x RSB e nang le Tween 20
• MilliQ metsi
• 2x Microcentrifuge tubes (1.5 mL)
• Pipette ka letsoho (10, 100 le 1000 μL)
• Sefa malebela a pipette

Pele o qala

• Qhibiliha Lib Tube haeba e hoamisitsoe.
• Qhibiliha RSB e leqhoa le Tween 20 mocheso oa kamore ka metsotso e 10. Boloka ho +4 °C ho fihlela o sebelisoa.
• Tšoaea li-tubes tse peli tse ncha tsa 1.5 mL microcentrifuge ka tsela e latelang:
  • Tšoaea tube e le 'ngoe "Dil" (bakeng sa laeborari e hlapolotsoeng ea 1:100)
  • Tšoaea tube e le 'ngoe "Seq" (bakeng sa ho itokisetsa ho kenya Laebrari)

Litaelo

1. Kenya 495 μL ea metsi a MilliQ ho Dil Tube.
2. Vortex the Lib Tube 'me ue sothe ka nakoana.
3. Ka bouena fetisetsa 5 μL ho tloha Lib Tube ho ea ho Dil Tube.
4. Vortex the Dil Tube 'me ue sothe ka nakoana.
5. Kenya 20 μL ea RBS ka Tween 20 ho Seq Tube.
6. Ka letsoho fetisetsa 20 μL ho tloha Dil Tube ho ea Seq Tube.
7. Vortex the Seq Tube 'me ue sothe ka nakoana.
8. Hang-hang tsoela pele ho 2.5 Load flow cell le Olink® Library ka har'a cartridge ea reagent.

HLOKOMELA: Boloka Lib Tube ho -20 °C haeba ho ka ba le liphetolelo hape.

Kenya phallo cell le Olink® Library ka har'a cartridge ea reagent

Nakong ea mohato ona, sele ea phallo le Laebrari ea Olink e hlapolotsoeng li kenngoa ka har'a cartridge ea reagent e qhibilihisitsoeng.

Lokisetsa benche

• 1x e qhibilihisitsoeng NextSeq™ 1000/2000 P2 Reagent Cartridge (100 cycles), e lokiselitsoeng mohato o fetileng
• 1x NextSeq™ 1000/2000 P2 Flow Cell, e lokiselitsoeng mohatong o fetileng.
• Seq Tube (e ikemiselitseng ho kenya Laeborari ea Olink e hlapolotsoeng), e lokiselitsoeng mohato o fetileng
• Pipette ka letsoho (100 μL)
• Pipette keletso (1 mL)

Lokisetsa khatriji

1. Tlosa cartridge ho tloha mokotleng oa silevera oa foil.
2. Fetola khatriji ka makhetlo a leshome ho kopanya ka botlalo li-reagents tse qhibilihisitsoeng ka hare.

HLOKOMELA: Ke ntho e tloaelehileng ho utloa likarolo tsa ka hare li lla.

Kenya sele e phallang ka har'a cartridge

1. Ha u se u itokiselitse ho kenya sele ea phallo ka har'a cartridge, tlosa sele ea phallo ho tloha sephutheloana. Tšoara sele ea phallo ho thebo ea bohlooho, leibole e shebileng holimo. Sebelisa liatlana tse sa lefelloeng tsa phofo ho qoba ho silafatsa bokaholimo ba khalase ea sele ea phallo.
2. Kenya sele ea phallo ka har'a sekotjana sa sele sa phallo se ka pele ho khatriji. Ho tobetsa ho utloahala ho bontša hore sele ea phallo e behiloe ka nepo.
3. Tlosa tab ea bohlooho ka ho e ntša.

Kenya Olink® Library ka har'a cartridge

1. Hlakola polokelo ea polokelo ea libuka ka ntlha e hloekileng ea 1 mL pipette.
2. Kenya 20 μL ea Laeborari ea Olink ho tloha Seq Tube ho ea tlase ho polokelo ea laeborari.

Etsa ts'ebetso ea tatellano ea Olink®

Nakong ea mohato ona, cartridge ea Buffer e nang le sele ea phallo e laetsoeng le Olink Library e kenngoa ho NextSeq™ 2000, 'me tatellano ea tatellano e qala ho sebelisoa recipe ea Olink.

Lokisetsa benche

• 1x NextSeq™ 1000/2000 P2 Reagent Cartridge (lipotoloho tse 100) e kentsoeng ka NextSeq™ 1000/2000 P2 Flow Cell le Olink Library e hlapolotsoeng, e lokiselitsoeng mohatong o fetileng.

Lokisa mokhoa oa ho matha

1. Ho tsoa ho menu ea software ea taolo, khetha Li-setting.
2. Tlas'a BaseSpace Sequence Hub Services & Proactive Support, khetha Local Run Setup.
3. Khetha Proactive Support Feela joalo ka li-setting tse ling. Ts'ebetso ena e hloka khokahano ea inthanete.
4. Khetha Sebaka sa Hosting bakeng sa data ea hau. Sebaka sa ho amohela se lokela ho ba haufi kapa haufi le sebaka sa heno.
5. Beha sebaka sa Output Folder bakeng sa data e tala ea hajoale. Kgetha Kgetha ho tsamaya ebe o kgetha foldara ya dipholo.
6. Khetha lebokose la "Denature le Dilute On Board" hore o ikemisetse le ho hlapolla Laeborari e ka har'a sesebelisoa.
7. Khetha lebokose la tlhahlobo la "Purge Reagent Cartridge" hore u tlose li-reagents tse sa sebelisoeng ka bohona karolong e sebelisitsoeng ea cartridge.
8. Khetha lebokose la tlhahlobo la Autocheck bakeng sa liapdeite tsa software ho itlhahloba ka boits'oaro bakeng sa liapdeite tsa software (ka boikhethelo). Ts'ebetso ena e hloka khokahano ea inthanete.
9. Kgetha Boloka.

Beha li-parameter tsa ho sebetsa

HLOKOMELA: Taelo ena e sebetsa ho mofuta oa 1.4 oa software ea taolo ea NextSeq™ 1000/2000. Tse ling tsa mehato e hlalositsoeng ka tlase e kanna ea fapana ha u sebelisa mofuta oa v1.2

1. Ho tsoa ho menu ea software ea taolo, khetha Qala.
2. Kgetha ka bowena Seta New Run ebe o tobetsa Seta.
3. Leqepheng la Run Setup, theha li-parameter tsa ho matha ka tsela e latelang:
   • Lefapheng la Run Name, kenya ID ea liteko e ikhethang.
   • Lethathamong la ho theoha ea Read Type, khetha khetho ea Single Read.
   • Kenya palo ea lipotoloho ka tsela e latelang:
     • Bala 1: 24
     • Index 1: 0
     • Index 2: 0
     • Bala 2: 0

HLOKOMELA: Ho bohlokoa hore Read 1 e behoe ho 24, ho seng joalo lebelo lohle le tla hloleha. Hlokomoloha melaetsa ea temoso ha u kenya palo ea lipotoloho.

• Lethathamong le theohang la Custom Primer Wells, khetha No.
• Lefapheng la Recipe ea Tloaelo (ea boikhethelo), khetha Khetha ho tsamaea ebe u khetha mokhoa o tloaelehileng oa recipe XML file Olink_NSQ2K_P2_V1. Kgetha Bula.
• Se ke oa kenya Sample Sheet.
• Etsa bonnete ba hore sebaka sa Output Folder se nepahetse. Ho seng joalo, khetha Khetha ho tsamaea ebe u khetha sebaka sa foldara ea tlhahiso eo u e batlang.
• Lebaleng la Denature le Dilute Onboard, khetha E nolofalitsoe ho tsoa lethathamong le theoha.
• Kgetha Prep.

Kenya khatriji e kentsoeng

1. Kgetha Mojaro. Visor ea sesebelisoa ea bula 'me terei ea ntšoa.
2. Beha khatriji e laetsoeng holim'a terei e shebileng holimo le sele ea phallo kahare ho sesebelisoa.
3. Kgetha Koala.
4. Hang ha cartridge e laetsoe hantle, netefatsa li-parameter tsa ho matha ebe u khetha Sequence. Sesebelisoa se etsa licheke tsa pele bakeng sa sesebelisoa le mokelikeli.
   • HLOKOMELA: Nakong ea tlhahlobo ea fluidics, ho lebelletsoe ho utloa melumo e mengata e hlahang.
5. Etsa bonnete ba hore ho matha ho qala ka mor'a hore tlhahlobo ea pele ea ho sebetsa e phethetsoe (~ metsotso e 15). Nako ea ho etsa tatellano ke hoo e ka bang 10h30 min.
   • HLOKOMELA: Bakeng sa liphoso leha e le life tsa ho hlahloba esale pele, sheba litaelo tsa moetsi. Hlokomela hore u se ke ua thulana kapa ua senya NextSeq™ 2000 nakong ea tatellano. Sesebelisoa sena se na le maikutlo a ho thothomela.
6. Hloekisa sebaka sa mosebetsi.

Lekola Tsoelo-pele ea Matha

Olink e sebelisa NGS joalo ka ho bala ho lekanya palo ea tatellano e tsebahalang e le ho hakanya bongata ba protheine e fanoeng ho s.amples (e amanang le tse ling tseamplets). Boleng ba data ho tsoa ho e 'ngoe le e' ngoe ea Explore sequencing run e khethoa haholo ke liparamente tsa QC tse ikhethang ho theknoloji ea Olink. Kahoo, metrics e tloaelehileng ea taolo ea boleng e sebelisoang ho NGS e tloaelehileng, joalo ka Q-score, ha e bohlokoa haholo.

Tlosa 'me u lahle cartridge ka mor'a ho matha

TLHOKOMELISO: Sehlopha sena sa li-reagents se na le lik'hemik'hale tse ka bang kotsi. Apara lisebelisoa tse lekaneng tsa ts'ireletso 'me u lahle li-reagents tse sebelisitsoeng ho latela litekanyetso tse sebetsang. Bakeng sa tlhaiso-leseling e batsi, sheba ho Illumina NextSeq 1000 le 2000 System Guide (tokomane #1000000109376).

1. Ha ts'ebetso e felile, khetha Eject Cartridge.
   • HLOKOMELA: Cartridge e sebelisitsoeng e kenyelletsang phallo cell e ka siuoa sebakeng ho fihlela e matha e latelang, empa eseng ho feta matsatsi a 3.
2. Tlosa cartridge ho tloha tereing.
3. Lahla li-reagents ho latela litekanyetso tse sebetsang.
4. Kgetha Koala Monyako. Terei e kentsoe hape.
5. Kgetha Lehae ho kgutlela ho Skrine sa lapeng.
   • HLOKOMELA: Kaha cartridge e na le mekhoa eohle ea ho tsamaisa tsamaiso, hammoho le letamo la ho bokella li-reagents tse sebelisitsoeng, ha ho hlokahale ho hlatsoa lisebelisoa ka mor'a ho matha.

Nalane ea ntlafatso

Phetolelo	Letsatsi	Tlhaloso
1.0	2021-12-01	E ncha

www.olink.com

Bakeng sa Tšebeliso ea Lipatlisiso Feela. Ha E sebelisoe Mekhoeng ea Tlhahlobo.

Sehlahisoa sena se kenyelletsa laesense ea tšebeliso e seng ea khoebo ea lihlahisoa tsa Olink. Basebelisi ba khoebo ba ka hloka lilaesense tse ling. Ka kopo ikopanye le Olink Proteomics AB bakeng sa lintlha. Ha ho na litiisetso, tse hlahisitsoeng kapa tse boletsoeng, tse fetang tlhaloso ena. Olink Proteomics AB ha e ikarabelle bakeng sa tšenyo ea thepa, kotsi ea motho, kapa tahlehelo ea moruo e bakiloeng ke sehlahisoa sena. Letšoao le latelang la khoebo ke la Olink Proteomics AB: Olink®. Sehlahisoa sena se koahetsoe ke litokelo tse 'maloa le likopo tsa patent tse fumanehang ho https://www.olink.com/patents/.

© Copyright 2021 Olink Proteomics AB. Matshwao ohle a kgwebo a mokga wa boraro ke thepa ya beng ba ona.

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

1193, v1.0, 2021-12-01

Litokomane / Lisebelisoa

Olink NextSeq 2000 Explore Sequencing System [pdf] Bukana ea Mosebelisi NextSeq 2000, Explore Sequencing System, NextSeq 2000 Explore Sequencing System

Litšupiso

• Bukana ea Mosebelisi