Olink NextSeq 550 Hlahloba Tatelano

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™ 550. 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 buka ea Olink® NPX Explore User Manual. 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 mokhoa oa ho hlophisa Lilaebrari tsa Olink ho NextSeq™ 550 u sebelisa NextSeq™ 500/550 High Output Kit v2.5 (75 Cycles). Protocol e sebelisetsoang ho latellana ke ho ikamahanya le Illumina® standard NGS workflow bakeng sa Illumina® NextSeq™ 550. Pele u tsoela pele ka ho latellana, etsa bonnete ba hore boleng ba Laebrari e 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 selefouno ea NextSeq™ 550 High Output flow le ho matha ka ngoe. Palo ea lisele tsa High Output flow tse hlokahalang ho tatellano ea mefuta e fapaneng ea Olink Explore Reagent Kits e hlalositsoe ho Lethathamo la 1. Haeba ho hlokahala ho matha ho fetang bonngoe, 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®

Nakong ea mohato ona, recipe ea tloaelo ea Olink® e kenngoa ho NextSeq™ 550. Mohato ona o hloka ho etsoa hanngoe feela, pele ho tsamaisoa ha tatellano ea Olink ho etsoa ka lekhetlo la pele.

HLOKOMELA: Recipe ea tloaelo ea Olink e tla sebetsa feela ka NextSeq™ 500/550 High Output Kits le NextSeq™ Control Software 4.0.

1. Unzip 'me u behe risepe ea tloaelo ea Olink Olink_NSQ550_HighOutput_V1 foldareng e latelang ea sesebelisoa sa NextSeq™ 550: C:\Program Files\Illumina\NextSeq Control Software\Recipe\Custom\High\.
2. Tlas'a Tsamaiso ea Sistimi> Laola Sesebelisoa, nolofalletsa Mekhoa ea Boipheliso. Haeba e sa khethiloe, khetho ea recipe e tloaelehileng e ke ke ea hlaha nakong ea ho seta.

HLOKOMELA

• Ho NCS 4.0 software version, khetho ea ho khetha recipe e tloaelehileng e tla etsahala feela ka mor'a hore cartridge ea reagent e kenngoe, eseng leqepheng la pele la ho seta.
• Ho matha ho tlameha ho behoa ka mokhoa oa matsoho ho lumella litlolo tse tloaelehileng.

Lokisetsa tatellano ea li-reagents

Nakong ea mohato ona, cartridge ea reagent e nang le li-clustering le li-reagents tsa ho latellana e qhibiliha 'me sele e phallang e lokisoa.

Lokisetsa khatriji ea reagent

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 550 System Guide (tokomane #15069765).

Lokisetsa benche

• 1x NextSeq™ 500/550 High Output Reagent Cartridge v2 (75 cycles).

Litaelo

1. Beha khatriji e nang le serame e qoelitsoeng halofo ea metsi a futhumetseng ka kamoreng 'me u e tlohele e qhibilihe hora e le 1. Etsa bonnete ba hore libaka tsohle tsa polokelo ea li-cartridges li qhibilihile ka ho feletseng.
   • HLOKOMELA: Bakeng sa boiketlo, qhibilihisa khatriji letsatsi pele 'me u e boloke bosiu ho 4 °C. Ka mocheso ona, li-reagents li tsitsitse ho fihlela beke e le 'ngoe.
2. Omisa setsi sa khatriji hantle ka thaole ea pampiri 'me u hlakole litiiso tse omeletseng ka lesela le se nang lesela ha ho hlokahala.
3. Fetola khatriji ka makhetlo a leshome ho kopanya ka botlalo li-reagents tse qhibilihisitsoeng ka hare.
4. Tlanya khatriji ka bonolo bencheng ho tlosa li-bubble tsa moea. Boloka cartridge ka mocheso oa kamore haeba e tla sebelisoa nakong ea lihora tse 4.

Lokisetsa sele ea phallo

Lokisetsa benche

• 1x NextSeq™ 500/550 High Output Flow Cell v2.5.

Litaelo

1. Tlisa sehatsetsing se phallang ka mocheso oa kamore bakeng sa metsotso e 30.
2. Apara liatlana tsa mahala tsa phofo (ho qoba ho silafatsa bokaholimo ba khalase ea sele ea phallo).
3. Ha u se u itokiselitse ho kenya sele ea phallo ka har'a sesebelisoa, tlosa sele ea phallo ho tloha sephutheloana le clamshell ea polasetiki.
4. Hlahloba sele e phallang. Haeba likaroloana kapa lerōle le bonahala holim'a leha e le efe ea khalase, hloekisa sebaka se sebetsang ka sesepa sa joala sa isopropyl se se nang lint 'me u se omise ka lab e nang le lesela le tlaase.

Lokisetsa Olink® Library bakeng sa tatellano

Nakong ea mohato ona, li-dilutions tsa NaOH le Tris-HCl lia lokisoa, 'me Olink Library e hloekisitsoeng le e laoloang ke boleng e hlapolloa le ho hlakisoa ka mehato e latellanang.

Lokisetsa NaOH dilution

• Ho hlapolla ha NaOH ho sebelisoa ho hlakisa Lilaebrari.

Lokisetsa benche

• 1 N NaOH stock
• MilliQ metsi
• 1x Microcentrifuge tube (1.5 mL)
• Pipette ka letsoho (10–100 μL)
• Sefa malebela a pipette

Pele o qala

• Tšoaea tube ea microcentrifuge "0.2 N NaOH".

Litaelo

1. Lokisetsa hlapo ea 0.2 N NaOH ka har'a Tube ea 0.2 N NaOH ho latela Lethathamo la 2.
2. Vortex the 0.2 N NaOH Tube hantle 'me u e sothe. Sebelisa nakong ea lihora tse 12.

Lethathamo la 2. 0.2 N NaOH dilution

Reagent	Bolumo (μL)
MilliQ metsi	80
1 N NaOH stock	20

Lokisetsa Tris-HCl dilution

• Thibelo ea Tris-HCl e sebelisoa ho nyenyefatsa Laeborari e sirelelitsoeng.

Lokisetsa benche

• 1 M Tris-HCl pH 7.0 stock (Trizma® hydrochloride solution)
• MilliQ metsi
• 1x Microcentrifuge tube (1.5 mL)
• Pipette ka letsoho (10–100 μL)
• Sefa malebela a pipette

Pele o qala

• Tšoaea tube ea microcentrifuge "Tris-HCl"

Litaelo

1. Lokisetsa 200 mM Tris-HCl ka har'a Tube ea Tris-HCl ho latela Lethathamo la 3.
2. Tlosa Tube ea Tris-HCl hantle 'me u e bilikise fatše.

Letlapa la 3. 200 mM Tris-HCl dilution

Reagent	Bolumo (μL)
MilliQ metsi	80
1M Tris-HCl pH 7.0 stock (Trizma® hydrochloride solution)	20

Hlakola Lilaebrari tsa Olink®

• Nakong ea mohato ona, Laebrari ea Olink e hloekisitsoeng le e laoloang ea boleng e hlapolotsoe 1:33.

Lokisetsa benche

• Lib Tube, e lokiselitsoeng ho latela Bukana e sebetsang ea Olink Explore User
• MilliQ metsi
• 1x Microcentrifuge tube (1.5 mL)
• Li-pipette tsa letsoho (0.5-10 le 100-1000 μL)
• Sefa malebela a pipette

Pele o qala

• Qhibiliha Lib Tube haeba e hoamisitsoe.
• Tšoaea tube e ncha ea microcentrifuge: "Dil".

Litaelo

1. Kenya 96 μL ea metsi a MilliQ ho Dil Tube.
2. Vortex the Lib Tube 'me ue sothe ka nakoana.
3. Fetisetsa 3 μL ho tloha Lib Tube ho ea ho Dil Tube.
4. Vortex the Dil Tube 'me ue sothe ka nakoana.

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

Hlakola 'me u hlapolle Laeborari ea Olink® sebakeng sa ho qetela sa ho kenya

Nakong ea mohato ona, Laeborari ea Olink e hlapollotsoeng e hlakotsoe 'me e hlapolotsoe ho fihlela mahloriso a ho qetela.

Lokisetsa benche

• Dil Tube, e lokiselitsoeng mohato o fetileng
• 0.2 N NaOH dilution, e sa tsoa lokisoa mohatong o fetileng
• 200 mM Tris-HCl (pH 7.0) dilution, e lokiselitsoeng mohatong o fetileng
• Hybridization buffer 1 (HT1) e kenyellelitsoe ka har'a NextSeq™ Accessory Box v2
• 2x Microcentrifuge tubes (1.5 mL le 2 mL)
• Li-pipette tsa letsoho (0.5-10 le 100-1000 μL)
• Sefa malebela a pipette

Pele o qala

• Qhibiliha buffer e hatselitsoeng ea HT1 mochesong oa kamore. Boloka ho +4 °C ho fihlela o sebelisoa.
• Tšoaea tšupu e ncha ea 1.5 mL microcentrifuge: "Den" (bakeng sa Laeborari e hloekisitsoeng).
• Tšoaea tube e ncha ea 2 mL microcentrifuge: "Seq" (bakeng sa ho itokisetsa ho kenya Library).

Litaelo

1. Fetisetsa 5 μL ho tloha ho Dil Tube ho ea ho Den Tube.
2. Kenya 5 μL ea 0.2 N NaOH ho Den Tube.
3. Vortex the Den Tube 'me ue sothe ka nakoana.
4. Eketsa Den Tube metsotso e 5 mocheso oa kamore ho fetola Laeborari.
5. Kenya 5 μL ea 200 mM Tris-HCl (pH 7.0) ho Den Tube ho fokotsa maikutlo.
6. Vortex the Den Tube 'me ue sothe ka nakoana.
7. Kenya 985 μL ea HT1 e chesitsoeng pele ho Den Tube.
8. Vortex the Den Tube 'me ue sothe ka nakoana. Tube e ka bolokoa ho +4 ° C ho fihlela e sebelisoa (letsatsing lona leo).
9. Fetisetsa 205 μL ho tloha Den Tube ho ea Seq Tube.
10. Kenya 1095 μL ea prechilled HT1 ho Seq Tube.
11. Fetolela Seq Tube ho kopanya li-reagents ebe o e bilika hanyane. Bophahamo ba ho qetela ba ho kenya ke 1.3 mL.
12. Hang-hang tsoela pele ho 2.5 Etsa ho latellana ha Olink®.

Etsa ts'ebetso ea tatellano ea Olink®

Nakong ea mohato ona, cartridge ea buffer, sele ea phallo le cartridge ea reagent e lokiselitsoeng e nang le Olink Library e kenngoa ho NextSeq 550, 'me ho latellana ho qala ho sebelisoa recipe ea tloaelo ea Olink.

Lokisetsa benche

• Seq Tube (e ikemiselitseng ho kenya Laebrari), e lokiselitsoeng mohato o fetileng
• 1x NextSeq™ 500/550 High Output Reagent Cartridge v2, e lokiselitsoeng mohato o fetileng
• 1x NextSeq™ 500/550 High Output Flow Cell v2.5, e lokiselitsoeng mohatong o fetileng.
• 1x NextSeq™ 500/550 Buffer Cartridge v2 (lipotoloho tse 75), ka mocheso oa kamore

Beha li-parameter tsa sequencing run

Nakong ea mohato ona, ho khethoa li-parameter tsa tatellano ho NextSeq™ 550.

1. Ho Skrine sa Lehae sa NextSeq™ 550, khetha Teko.
2. Ho Skrine sa Select Assay, khetha Sequence.
3. Leqepheng la Run Setup, khetha "Manual run mode" ebe u E latelang.
4. Beha li-parameter tsa "run" ka tsela e latelang:
   • Lefapheng la Run Name, kenya ID ea liteko e ikhethang.
   • Lebaleng la ID ea Laebrari, kenya ID ea Laebrari eo u e tsamaisang (ka boikhethelo).
   • Lefapheng la Mofuta oa Bala, 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
     • BOHLOKOA: Ho bohlokoa hore Read 1 e behoe ho 24, ho seng joalo lebelo lohle le tla hloleha.
   • Boloka lebokose la ho hlahloba bakeng sa li-primer tsa tloaelo le sa khethoa.
   • Beha sebaka sa foldara ea tlhahiso bakeng sa data e tala ea hajoale. Kgetha Browse ho fetola sebaka sa foldara ya diphoho.
5. U se ke ua theha Sample Sheet.
6. Khetha Tlosa lisebelisoa bakeng sa ts'ebetso ena.
7. Khetha E 'ngoe.

Kenya phallo sele ho NextSeq™ 550

1. Tlosa sele e sebelisitsoeng ea phallo ho tloha nakong e fetileng.
2. Beha sele e ncha e lokiselitsoeng ea phallo ho stage.
3. Kgetha Mojaro. Lemati le koetsoe ka bo eona.
4. Ha "flow cell ID" e hlaha skrineng 'me li-sensor li hlahlojoa ka botala, khetha E latelang.

Tlosa setshelo sa reagent

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 Tataiso ea Sistimi ea Illumina NextSeq 550.

1. Bula lemati la phaposi ea buffer, tlosa sejana sa li-reagents tse sebelisitsoeng karolong e ka tlase, 'me u lahle litaba ho latela litekanyetso tse sebetsang.
2. Tsamaisa setshelo se se nang letho ho khutlela karolong e ka tlase ea buffer. Tobetsa e utlwahalang e bontsha hore setshelo se behilwe ka nepo.

Kenya khatriji ea buffer

1. Tlosa "buffer cartridge" e sebelisitsoeng karolong e kaholimo ea "buffer" 'me u lahle litaba ho latela maemo a sebetsang.
2. Tsamaisa khatriji e ncha ea buffer karolong e ka holimo ea buffer. Ho penya ka mokhoa o utloahalang ho bontša hore cartridge e behiloe ka nepo. Etsa bonnete ba hore ID ea buffer cartridge e hlaha skrineng le hore li-sensor li hlahlojoe li le tala.
3. Kwala lemati la buffer compartment ebe o kgetha E latelang.

Kenya khatriji ea reagent

1. Bula lemati la phaposi ea reagent, tlosa khatriji e sebelisoang, 'me u lahle lintho tse sa sebelisoeng ho latela maemo a sebetsang. Letamo le boemong ba 6 lea ntšoa ho thusa ho lahla ka mokhoa o sireletsehileng.
2. Hlakola tiiso ea letamo #10 e ngotsoeng "Laebrari ea Tlala Mona" ka ntlha e hloekileng ea 1 mL pipette.
3. Laela 1.3 mL Laeborari ea Olink ho tloha Seq Tube ho ea polokelong ea #10 e ngotsoeng "Laebrari ea Tlala Mona".
4. Tsamaisa cartridge e ncha ea reagent ka phaposing ea reagent ebe u koala lemati la phaposi ea reagent.
5. Khetha Mojaro 'me u eme metsotsoana ~ 30 ho fihlela ID ea cartridge ea reagent e hlaha skrineng mme li-sensor li hlahlojoa li le tala.
6. Ho tsoa lethathamong le theohang la Recipe, khetha khetho ea [Moetlo] "Olink_NSQ550_HighOutput_V1".
   • BOHLOKOA: Etsa bonnete ba hore risepe e tloaelehileng e ne e kentsoe sesebelisoa pele. Sheba 2.2 Kenya risepe ea tloaelo ea Olink®.
7. Khetha E 'ngoe.

1. Netefatsa li-parameter tse bonts'itsoeng ho Review skrineng. Ho hlophisa liparamente life kapa life, tobetsa Khutlela ho khutlela skrineng sa Run Setup.
2. Khetha E 'ngoe. Ho matha ho qala ka mor'a tlhahlobo ea othomathike esale pele. Nako ea ho etsa tatellano ke hoo e ka bang 7h30 min.
3. BOHLOKOA: Etsa bonnete ba hore ho matha ho qala ha cheke ea pele ea othomathiki e phethiloe (~ metsotso e 5).
   1. HLOKOMELABakeng sa liphoso leha e le life tsa ho hlahloba esale pele, sheba litaelo tsa moetsi.
   2. HLOKOMELA: Hlokomela hore u se ke ua thulana kapa ua senya NextSeq™ 550 nakong ea tatellano. Sesebelisoa sena se na le maikutlo a ho thothomela.
4. Hloekisa sebaka sa mosebetsi.
   1. HLOKOMELA: Ha ts'ebetso ea tatellano e phethiloe, software e qala ho itlhatsoa ka mor'a ho qeta ho sebelisa litharollo tsa ho hlatsoa tse fanoeng ka cartridge ea buffer le NaOCl e fanoeng ka cartridge ea reagent. Ho hlatsoa hona ho nka metsotso e ka bang 90. Konopo ea Lehae e qala ho sebetsa hang ha ho hlatsuoa ho felile. Li-cartridges tse sebelisitsoeng le sele ea phallo li ka siuoa sebakeng ho fihlela ho matha ho latelang.

Lekola tsoelo-pele ea ho 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.

Nalane ea ntlafatso

Phetolelo	Letsatsi	Tlhaloso
1.2	2022-11-01	1.2 E nkeloe sebaka ke Olink® Mydata ka Olink® NPX Explore. 2.4 E kentse 0.2 N ho Pele o qala.
1.1	2021-12-13	Liphetoho tsa bohlophisi
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 a kgwebo kaofela ke thepa ya beng ba ona.
Olink Proteomics, Dag Hammarskjölds väg 52B , SE-752 37 Uppsala, Sweden
1192, v1.2, 2022-11-01

Litokomane / Lisebelisoa

Olink NextSeq 550 Hlahloba Tatelano [pdf] Bukana ea Mosebelisi NextSeq 550 Hlahloba Tatelano, NextSeq 550, Hlahloba Tatelano, Tatelano

Litšupiso

• Bukana ea Mosebelisi