Olink NextSeq 2000 Bincika Tsarin Bincika

Bayanan kula

Littafin Jagorar Mai amfani na Olink®, doc nr 1153, ya ƙare, kuma an maye gurbinsu da waɗannan takaddun:

• Olink® Explore Overview Littafin mai amfani, doc nr 1187
• Olink® Explore 384 Jagoran mai amfani, doc nr 1188
• Olink® Explore 4 x 384 Jagoran mai amfani, doc nr 1189
• Olink® Explore 1536 & Fadada Jagorar Mai Amfani, Doc nr 1190
• Olink® Explore 3072 Jagoran mai amfani, doc nr 1191
• Olink® Binciko Jeri ta amfani da NextSeq 550 Manual User, doc nr 1192
• Olink® Binciko Jeri ta amfani da NextSeq 2000 Manual User, doc nr 1193
• Olink® Binciko Jeri ta amfani da NovaSeq 6000 Manual User, doc nr 1194

Gabatarwa

Amfani da niyya

Olink® Explore dandamali ne na immunoassay na multix don gano furotin na ɗan adam. An yi nufin samfurin don Amfani da Bincike kawai, kuma ba don amfani da shi ba a hanyoyin bincike. Kwararrun ma'aikatan dakin gwaje-gwaje ne kawai za su gudanar da aikin dakin gwaje-gwaje. ƙwararrun ma'aikata ne kawai za a gudanar da sarrafa bayanai. Sakamakon ana nufin masu bincike su yi amfani da su tare da wasu binciken asibiti ko dakin gwaje-gwaje.

Game da wannan littafin

Wannan Jagorar Mai Amfani yana ba da umarnin da ake buƙata don jera Olink® Explore Laburaren a kan Illumina® NextSeq™ 2000. Dole ne a bi umarnin sosai kuma a bayyane. Duk wani sabani a cikin matakan dakin gwaje-gwaje na iya haifar da nakasa bayanai. Kafin fara aikin dakin gwaje-gwaje, tuntuɓi Olink® Explore Overview Jagorar mai amfani don gabatarwa ga dandamali, gami da bayani game da reagents, kayan aiki da takaddun da ake buƙata, ƙarewaview na tafiyar aiki, da kuma jagororin dakin gwaje-gwaje. Don umarni kan yadda ake gudanar da Olink® Explore Reagent Kits, koma zuwa Manual mai amfani na Olink® Explore. Don sarrafa bayanai da bincike na sakamakon Olink® Explore jerin sakamakon, koma zuwa Jagorar Mai amfani na Olink® MyData Cloud. Duk alamun kasuwanci da haƙƙin mallaka da ke ƙunshe a cikin wannan kayan mallakar Olink® Proteomics AB ne, sai dai in an faɗi.

Goyon bayan sana'a

Don tallafin fasaha, tuntuɓi Olink Proteomics a: support@olink.com.

Umurnin dakin gwaje-gwaje

Wannan babin yana ba da umarni kan yadda ake jera Laburaren Olink akan NextSeq™ 2000 ta amfani da NextSeq™ 1000/2000 P2 Reagents (100 Cycles) v3. Yarjejeniyar da aka yi amfani da ita don tsarawa shine daidaitawa na Illumina® daidaitaccen aikin NGS don Illumina® NextSeq ™ 2000. Kafin a ci gaba zuwa jerin, tabbatar cewa an tabbatar da ingancin Laburaren Olink mai tsabta. Koma zuwa aikin Olink Explore Manual mai amfani don umarni game da sarrafa inganci.

Tsara tsarin gudu

Ana iya jera Laburaren Olink ɗaya ta hanyar tantanin halitta mai gudana na NextSeq™ 2000 P2 da kowane gudu. Adadin sel masu kwarara na P2 da gudanar da ake buƙata don jeri daban-daban Olink Explore Reagent Kits an kwatanta su a cikin Tebur 1. Idan ana buƙatar gudu fiye da ɗaya, maimaita umarnin da aka bayyana a cikin wannan jagorar.

Tebura 1. Tsare-tsare na gudu

Olink® Explore Reagent Kit	Adadin Laburaren Olink	Adadin tantanin halitta (s) da run(s)
Olink® Explore 384 Reagent Kit	1	1
Olink® Binciken 4 x 384 Reagent Kit	4	4
Olink® Explore 1536 Reagent Kit	4	4
Olink® Binciko Fadada Reagent Kit	4	4
Olink® Explore 3072 Reagent Kit	8	8

Shigar Olink® girke-girke na al'ada

Ajiye girke-girke na Olink na al'ada xml-file Olink_NSQ2K_P2_V1 a cikin babban fayil ɗin kayan aiki da ya dace.

NOTE: Girke-girke na al'ada na Olink zai yi aiki ne kawai tare da NextSeq ™ 1000/2000 P2 Reagents (100 Cycles) v3 kit da NextSeq ™ 1000/2000 software mai sarrafawa v1.2 ko v1.4.

Shirya jerin reagents

A lokacin wannan matakin, reagent harsashi mai ƙunshe da tari da reagents yana narke kuma ana shirya tantanin da ke gudana.

GARGADI: Harsashin reagent ya ƙunshi wasu sinadarai masu haɗari. Saka isassun kayan kariya kuma a zubar da reagents da aka yi amfani da su daidai da ma'auni masu dacewa. Don ƙarin bayani, koma zuwa Illumina NextSeq 1000 da Jagoran Tsarin 2000 (takardun #1000000109376).

Shirya reagent harsashi

Ana iya narke harsashin da ba a buɗe ba ta amfani da hanyoyi daban-daban guda uku: a dakin da zafin jiki, a cikin ruwan wanka mai sarrafawa, ko a cikin firiji.

Shirya benci

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

Umarni

1. Narke harsashin reagent kamar yadda aka bayyana a cikin Table 2.

Table 2. Reagent harsashi narke hanyoyin

Hanyar narkewa	Umarni
A dakin da zafin jiki	Ba tare da buɗe jakar jakar azurfa ba, sanya daskararre reagent a kan benci kuma a narke shi a cikin ɗaki na tsawon awanni 9. Kada ku wuce awanni 16. Tabbatar cewa alamar jakar tana fuskantar sama kuma iska zata iya yawo a kusa da harsashi.
A cikin ruwan wanka	Ba tare da buɗe jakar foil ɗin azurfa ba, sanya kwandon reagent ɗin daskararre a cikin ruwan wanka mai ƙarfi na 25 ° C kuma bar shi ya narke na tsawon awanni 6. Kada ku wuce awa 8. Tabbatar cewa alamar jakar tana fuskantar sama kuma harsashi baya juyawa yayin narke. A bushe harsashi sosai tare da tawul na takarda.
A cikin firiji	Ba tare da buɗe jakar jakar azurfa ba, sanya harsashi a kan benci kuma a narke shi a dakin da zafin jiki na tsawon awanni 6. Tabbatar cewa alamar jakar tana fuskantar sama kuma iska zata iya yawo a kusa da harsashi. Gama narke harsashi a cikin firiji na tsawon awanni 12. Kada ku wuce awanni 72. Kafin gudu, cire kwas ɗin da ba a buɗe ba daga firiji kuma ba shi damar isa ga zafin jiki a cikin akalla minti 15, amma ba fiye da awa 1 ba.

NOTE: Ba za a iya sake daskarar da harsashin narke ba kuma dole ne a adana su a zafin jiki na 4 ° C, na tsawon sa'o'i 72.

Shirya tantanin halitta kwarara

Shirya benci

• 1x NextSeq™ 1000/2000 P2 Flow Cell

Umarni

1. Kawo tantanin da ke kwarara cikin firiji zuwa zafin jiki na minti 10-15.

Shirya Laburaren Olink® don jeri

A yayin wannan matakin, ana diluted da tsarkakewa da ingancin Laburaren Olink zuwa babban taro na ƙarshe. Lura cewa denaturation Library ana yin ta atomatik a kan kayan aiki.

Shirya benci

• Lib Tube, wanda aka shirya bisa ga umarnin mai amfani na Olink Explore
• 1 x RSB tare da Tween 20
• Ruwan MilliQ
• 2 x Microcentrifuge bututu (1.5 ml)
• pipette na hannu (10, 100 da 1000 μL)
• Tace tukwici

Kafin ka fara

• Narke Lib Tube idan ya daskare.
• Narke daskararre RSB tare da Tween 20 a dakin da zafin jiki na minti 10. Ajiye a +4 ° C har sai amfani.
• Alama sababbin bututun microcentrifuge 1.5 ml kamar haka:
  • Alama bututu ɗaya "Dil" (don 1:100 diluted Library)
  • Alama bututu ɗaya "Seq" (don shirye-shiryen loda ɗakin karatu)

Umarni

1. Ƙara 495 ​​μL na Ruwan MilliQ zuwa Dil Tube.
2. Vortex da Lib Tube kuma juya shi a takaice.
3. Canja wurin 5 μL da hannu daga Lib Tube zuwa Dil Tube.
4. Vortex da Dil Tube kuma juya shi a takaice.
5. Ƙara 20 μL na RBS tare da Tween 20 zuwa Seq Tube.
6. Canja wurin 20 μL da hannu daga Dil Tube zuwa Seq Tube.
7. Vortex da Seq Tube kuma juya shi a takaice.
8. Nan da nan ci gaba zuwa 2.5 Load flow cell da Olink® Library cikin reagent harsashi.

NOTE: Ajiye Tube(s) na Lib a -20 °C idan akwai yiwuwar sake kunnawa.

Load cell flow cell da Olink® Library cikin reagent harsashi

A lokacin wannan matakin, ana loda tantanin halitta mai gudana da Laburaren Olink da aka diluted a cikin narkar da reagent.

Shirya benci

• 1x narke NextSeq ™ 1000/2000 P2 Reagent Cartridge (100 hawan keke), wanda aka shirya a mataki na baya
• 1x NextSeq™ 1000/2000 P2 Flow Cell, wanda aka shirya a mataki na baya
• Seq Tube (tare da shirye don loda diluted Olink Library), wanda aka shirya a mataki na baya
• pipette na hannu (100 μL)
• Tushen pipette (1 ml)

Shirya harsashi

1. Cire harsashi daga jakar foil na azurfa.
2. Juya harsashi sau goma don haɗawa da narke reagents sosai a ciki.

NOTE: Yana da al'ada don jin jingling abubuwan haɗin ciki.

Loda tantanin halitta mai gudana a cikin katun

1. Lokacin da aka shirya don loda tantanin halitta mai gudana a cikin katun, cire tantanin halitta mai gudana daga kunshin. Riƙe cell ɗin da ke gudana ta shafin launin toka, tare da lakabin akan shafin yana fuskantar sama. Yi amfani da safofin hannu marasa foda don gujewa gurɓata saman gilashin tantanin halitta.
2. Saka tantanin halitta mai gudana a cikin ramin kwayar tantanin halitta a gaban harsashi. Dannawa mai ji yana nuna cewa an sanya tantanin halitta mai gudana daidai.
3. Cire shafin launin toka ta hanyar ciro shi.

Loda ɗakin karatu na Olink a cikin katun

1. Soki tafki na Laburare tare da tsaftataccen tip 1 ml na pipette.
2. Load da 20 μL na Laburaren Olink daga Seq Tube zuwa kasan tafki na ɗakin karatu.

Yi Olink® gudu-gudu

A yayin wannan matakin, an ɗora buffer cartridge tare da ɗigon ruwa mai ɗorewa da kuma Olink Library a cikin NextSeq™ 2000, kuma ana fara aiwatar da tsarin ta amfani da girke-girke na Olink.

Shirya benci

• 1x NextSeq ™ 1000/2000 P2 Reagent Cartridge (100 cycles) wanda aka ɗora tare da NextSeq ™ 1000/2000 P2 Flow Cell da Laburaren Olink mai diluted, wanda aka shirya a mataki na baya.

Sanya Yanayin Gudu

1. Daga menu na sarrafa software, zaɓi Saituna.
2. Ƙarƙashin Sabis na BaseSpace Sequence Hub & Taimako Mai Sauƙi, zaɓi Saitin Gudun Gida.
3. Zaɓi Taimako Mai Haɓakawa Kawai azaman ƙarin saituna. Wannan aikin yana buƙatar haɗin intanet.
4. Zaɓi wurin Hosting don bayanan ku. Wurin Hosting yakamata ya kasance a ciki ko kusa da yankin ku.
5. Saita wurin babban fayil ɗin fitarwa don ɗanyen bayanai masu gudana na yanzu. Zaɓi Zaɓi don kewaya kuma zaɓi babban fayil ɗin fitarwa.
6. Zaɓi Akwatin rajistan Denature da Dilute On Board don cirewa ta atomatik kuma tsarma ɗakin ɗakin karatu da ke kan kayan aikin.
7. Zaɓi akwatin rajistan Reagent don share reagents ta atomatik zuwa ɓangaren reagents da aka kashe na harsashi.
8. Zaɓi akwatin rajistan Autocheck don sabunta software don bincika sabunta software ta atomatik (na zaɓi). Wannan aikin yana buƙatar haɗin intanet.
9. Zaɓi Ajiye.

Saita sigogin gudu

NOTE: Wannan umarnin ya shafi sigar 1.4 na software na sarrafa NextSeq™ 1000/2000. Wasu matakan da aka bayyana a ƙasa na iya bambanta yayin amfani da sigar v1.2

1. Daga menu na sarrafa software, zaɓi Fara.
2. Zaɓi Saita Sabon Run da hannu kuma danna Saita.
3. A cikin Run Setup, saita sigogin gudu kamar haka:
   • A cikin filin Run Sunan, shigar da ID ɗin gwaji na musamman.
   • A cikin jerin saukarwa Na Nau'in Karatu, zaɓi zaɓin Karatu ɗaya.
   • Shigar da adadin zagayowar kamar haka:
     • Karanta 1: 24
     • Fihirisa 1: 0
     • Fihirisa 2: 0
     • Karanta 2: 0

NOTE: Yana da mahimmanci cewa an saita karatun 1 zuwa 24, in ba haka ba duk gudu zai gaza. Yi watsi da saƙonnin gargaɗi lokacin shigar da adadin zagayowar.

• A cikin jerin zaɓuka na Custom Primer Wells, zaɓi A'a.
• A cikin filin girke-girke na al'ada (na zaɓi), zaɓi Zaɓi don kewaya kuma zaɓi girke-girke na al'ada XML file Olink_NSQ2K_P2_V1. Zaɓi Buɗe.
• Kar a shigo da Sampda Sheet.
• Tabbatar cewa wurin babban fayil ɗin fitarwa daidai ne. In ba haka ba, zaɓi Zaɓi don kewaya kuma zaɓi wurin babban fayil ɗin fitarwa da ake so.
• A cikin filin Denature da Dilute Onboard, zaɓi An kunna daga jerin zaɓuka.
• Zaɓi Prep.

Loda kwandon da aka ɗora

1. Zaɓi Load. Visor na kayan aiki yana buɗewa kuma aka fitar da tire.
2. Sanya harsashin da aka ɗora a kan tire tare da alamar tana fuskantar sama da tantanin da ke gudana a cikin kayan aiki.
3. Zaɓi Kusa.
4. Da zarar an ɗora nauyin harsashi da kyau, tabbatar da sigogin gudu kuma zaɓi Jeri. Kayan aikin yana yin pre-gudu cak don kayan aiki da masu ruwa.
   • NOTE: Yayin binciken ruwa, ana sa ran jin sautuna da dama.
5. Tabbatar cewa gudu yana farawa bayan an kammala bincike na atomatik (~ mintoci 15). Lokacin gudanar da jeri yana kusan 10h30 min.
   • NOTE: Ga kowane gazawar rajistan da aka riga aka yi, koma zuwa umarnin masana'anta. Yi hankali don kada ku shiga ciki ko in ba haka ba ku dame NextSeq™ 2000 yayin gudanar da jerin abubuwan. Kayan aiki yana kula da rawar jiki.
6. Tsaftace wurin aiki.

Saka idanu Gudun Ci gaba

Olink yana amfani da NGS azaman karantawa don ƙididdige adadin sanannun jerin don ƙididdige yawan adadin furotin da aka bayar a cikin s.amples (dangane da sauran samples). Ingancin bayanai daga kowane Bincika gudanar da jerin gwano an ƙaddara ta musamman ta sigogin QC na musamman ga fasahar Olink. Don haka, daidaitattun ma'aunin sarrafa ingancin da aka yi amfani da su a cikin NGS na al'ada, kamar Q-score, ba su da mahimmanci.

Fitar da jefar da harsashi bayan gudu

GARGADI: Wannan saitin reagents ya ƙunshi sinadarai masu haɗari masu haɗari. Saka isassun kayan kariya kuma a zubar da reagents da aka yi amfani da su daidai da ma'auni masu dacewa. Don ƙarin bayani, koma zuwa Illumina NextSeq 1000 da Jagoran Tsarin 2000 (takardun #1000000109376).

1. Lokacin da aikin ya cika, zaɓi Fitar da Cartridge.
   • NOTE: Harsashin da aka yi amfani da shi ciki har da tantanin halitta za a iya barin shi a wurin har sai na gaba, amma bai wuce kwanaki 3 ba.
2. Cire katun daga tire.
3. Zubar da reagents daidai da ma'auni masu dacewa.
4. Zaɓi Rufe Ƙofa. An sake loda tire.
5. Zaɓi Gida don komawa kan Fuskar allo.
   • NOTE: Tun da harsashi ya ƙunshi duk hanyoyin da za a gudanar da tsarin, da kuma tafki don tattara reagents da aka yi amfani da su, babu buƙatar wanke kayan aiki bayan gudu.

Tarihin bita

Sigar	Kwanan wata	Bayani
1.0	2021-12-01	Sabo

www.olink.com

Don Amfanin Bincike Kawai. Ba don Amfani ba a Tsarin Bincike.

Wannan samfurin ya haɗa da lasisi don amfanin samfuran Olink ba na kasuwanci ba. Masu amfani da kasuwanci na iya buƙatar ƙarin lasisi. Tuntuɓi Olink Proteomics AB don cikakkun bayanai. Babu wani garanti, bayyana ko fayyace, wanda ya wuce wannan bayanin. Olink Proteomics AB bashi da alhakin lalacewar dukiya, rauni na mutum, ko asarar tattalin arzikin da wannan samfurin ya haifar. Alamar kasuwanci mai zuwa mallakar Olink Proteomics AB: Olink®. Wannan samfurin yana rufe da haƙƙin mallaka da aikace-aikacen haƙƙin mallaka da ake samu a https://www.olink.com/patents/.

© Haƙƙin mallaka 2021 Olink Proteomics AB. Duk alamun kasuwanci na ɓangare na uku mallakin masu su ne.

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

1193, v1.0, 2021-12-01

Takardu / Albarkatu

Olink NextSeq 2000 Bincika Tsarin Bincika [pdf] Manual mai amfani Na gaba Seq 2000, Bincika Tsarin Bincika, Na gabaSeq 2000 Binciko Tsarin Tsarin

Magana

• Manual mai amfani