Olink NextSeq 550 Bincika Jeri
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™ 550. 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 nazarin sakamakon Olink® Explore jere, koma zuwa Olink® NPX Explore Manual User. 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 dakunan karatu na Olink akan NextSeq™ 550 ta amfani da NextSeq™ 500/550 High Output Kit v2.5 (75 Cycles). Yarjejeniyar da aka yi amfani da ita don tsarawa shine daidaitawa na Illumina® daidaitaccen aikin NGS don Illumina® NextSeq ™ 550. Kafin a ci gaba zuwa jerin, tabbatar cewa an tabbatar da ingancin ɗakin karatu mai tsabta. Koma zuwa aikace-aikacen Olink Explore Manual don umarni game da sarrafa inganci.
Tsara tsarin gudu
Ɗayan Laburaren Olink za a iya jera shi ta hanyar NextSeq™ 550 Babban Fitarwa mai gudana da kowane gudu. Adadin sel masu gudana mai girma da gudana da ake buƙata don jeri daban-daban Olink Explore Reagent Kits an kwatanta su a cikin Tebu 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
A lokacin wannan mataki, an shigar da girke-girke na al'ada na Olink® akan NextSeq™ 550. Wannan matakin yana buƙatar yin sau ɗaya kawai, kafin a gudanar da tsarin Olink a karon farko.
NOTE: Girke-girke na al'ada na Olink zai yi aiki ne kawai tare da NextSeq ™ 500/550 High Output Kits da NextSeq ™ Control Software 4.0.
- Cire zip ɗin kuma sanya girke-girke na Olink na al'ada Olink_NSQ550_HighOutput_V1 a cikin babban fayil mai zuwa na kayan aikin NextSeq™ 550: C:\Program Files\Ilumina\NextSeq Control SoftwareRecipeCustomHigh\.
- A karkashin tsarin tsari> Sarrafa kayan aiki, kunna girke-girke na al'ada. Idan ba a zaɓa ba, zaɓin girke-girke na al'ada ba zai bayyana yayin saitin gudu ba.
NOTE
- A cikin sigar software ta NCS 4.0, zaɓin zaɓin girke-girke na al'ada zai faru ne kawai bayan an ɗora kayan reagent, ba a cikin shafin saitin farko ba.
- Dole ne a saita gudu a yanayin jagora don ba da damar girke-girke na al'ada.
Shirya jerin reagents
A lokacin wannan matakin, reagent harsashi mai ƙunshe da tarin reagents yana narke kuma an shirya tantanin da ke gudana.
Shirya reagent harsashi
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 550 Jagoran Tsarin (takardun #15069765).
Shirya benci
- 1x NextSeq ™ 500/550 Babban Fitarwa Reagent Cartridge v2 (zagaye 75).
Umarni
- Sanya daskararre reagent cartridge rabin-zurfin cikin ruwa mai zafin daki kuma bar shi ya narke na awa 1. Tabbatar cewa duk reagent reservoirs na harsashi an narke gaba daya.
- NOTE: Don saukakawa, narke harsashi a ranar da ta gabata kuma adana shi cikin dare a 4 ° C. A wannan zafin jiki, reagents suna barga har zuwa mako guda.
- A busar da gindin harsashi sosai da tawul na takarda sannan a goge hatimin da ya bushe da abin da ba shi da lint idan an buƙata.
- Juya harsashi sau goma don haɗawa da narke reagents sosai a ciki.
- Matsa katun a hankali akan benci don cire kumfa. Ajiye katun a dakin da zafin jiki idan za a yi amfani da shi a cikin sa'o'i 4.
Shirya tantanin halitta kwarara
Shirya benci
- 1x NextSeq™ 500/550 Babban Fitowar Fitowa Mai Yawo 2.5.
Umarni
- Kawo tantanin dake kwarara cikin firiji zuwa zafin daki na tsawon mintuna 30.
- Saka sabon safofin hannu na foda (don guje wa gurɓata fuskar gilashin tantanin halitta).
- Lokacin da aka shirya don loda tantanin halitta mai gudana a cikin kayan aiki, cire tantanin halitta mai gudana daga fakitin da clamshell na filastik.
- Duba tantanin da ke gudana. Idan ƙura ko ƙura ta ganuwa akan kowane saman gilashin, tsaftace wurin da ake amfani da shi tare da gogewar isopropyl barasa mara lint kuma a bushe shi da ƙananan nama mai ƙarancin lint.
Shirya Laburaren Olink® don jeri
A yayin wannan matakin, ana shirya dilutions NaOH da Tris-HCl, kuma ana diluted Laburaren Olink mai tsabta da inganci kuma an cire shi cikin matakai masu zuwa.
Shirya NaOH dilution
- Ana amfani da dilution na NaOH don hana ɗakunan karatu.
Shirya benci
- 1 NNOH hannun jari
- Ruwan MilliQ
- 1 x Microcentrifuge tube (1.5 ml)
- pipette na hannu (10-100 μL)
- Tace tukwici
Kafin ka fara
- Alama bututun microcentrifuge "0.2 N NaOH".
Umarni
- Shirya dilution 0.2 N NaOH a cikin 0.2 N NaOH Tube bisa ga Table 2.
- Vortex da 0.2 N NaOH Tube sosai kuma ya juya ƙasa. Yi amfani a cikin sa'o'i 12.
Tebur 2. 0.2 N NaOH dilution
| Reagent | girma (μL) |
| Ruwan MilliQ | 80 |
| 1 NNOH hannun jari | 20 |
Shirya Tris-HCl dilution
- Ana amfani da dilution na Tris-HCl don kawar da Laburaren da aka ba da izini.
Shirya benci
- 1M Tris-HCl pH 7.0 stock (Trizma® hydrochloride bayani)
- Ruwan MilliQ
- 1 x Microcentrifuge tube (1.5 ml)
- pipette na hannu (10-100 μL)
- Tace tukwici
Kafin ka fara
- Alama bututun microcentrifuge "Tris-HCl"
Umarni
- Shirya 200mM Tris-HCl dilution a cikin Tris-HCl Tube bisa ga Table 3.
- Vortex da Tris-HCl Tube sosai kuma a jujjuya shi.
Tebur 3. 200 mM Tris-HCl dilution
| Reagent | girma (μL) |
| Ruwan MilliQ | 80 |
| 1M Tris-HCl pH 7.0 stock (Trizma® hydrochloride bayani) | 20 |
Dilute Olink® Library
- A yayin wannan matakin, ana diluted da ingantaccen ɗakin karatu na Olink mai inganci 1:33.
Shirya benci
- Lib Tube, wanda aka shirya bisa ga umarnin mai amfani na Olink Explore
- Ruwan MilliQ
- 1 x Microcentrifuge tube (1.5 ml)
- pipettes na hannu (0.5-10 da 100-1000 μL)
- Tace tukwici
Kafin ka fara
- Narke Lib Tube idan ya daskare.
- Alama sabon bututun microcentrifuge: “Dil”.
Umarni
- Ƙara 96 μL na Ruwan MilliQ zuwa Dil Tube.
- Vortex da Lib Tube kuma juya shi a takaice.
- Canja wurin 3 μL daga Lib Tube zuwa Dil Tube.
- Vortex da Dil Tube kuma juya shi a takaice.
NOTE: Ajiye Tube(s) na Lib a -20 °C idan akwai yiwuwar sake kunnawa.
Denature da tsoma Olink® Library zuwa karshe loading maida hankali
A yayin wannan matakin, ɗakin karatu na Olink ɗin da aka diluted yana hana shi kuma yana ƙara diluted zuwa maida hankali na ƙarshe.
Shirya benci
- Dil Tube, wanda aka shirya a mataki na baya
- 0.2 N NaOH dilution, sabon shiri a mataki na baya
- 200 mM Tris-HCl (pH 7.0) dilution, wanda aka shirya a mataki na baya
- Haɓakawa 1 (HT1) an haɗa shi a cikin Akwatin Naɗi na NextSeq 2
- 2 x Microcentrifuge bututu (1.5 ml da 2 ml)
- pipettes na hannu (0.5-10 da 100-1000 μL)
- Tace tukwici
Kafin ka fara
- Narke daskararrun buffer HT1 a zazzabi na ɗaki. Ajiye a +4 ° C har sai amfani.
- Alama sabon bututu microcentrifuge 1.5 ml: “Den” (na Laburaren da aka ba da izini).
- Alama sabon bututu microcentrifuge 2 ml: “Seq” (don shirye-shiryen loda Laburare).
Umarni
- Canja wurin 5 μL daga Dil Tube zuwa Den Tube.
- Ƙara 5 μL na 0.2 N NaOH zuwa Den Tube.
- Vortex da Den Tube kuma juya shi a takaice.
- Ƙaddamar da Den Tube na tsawon mintuna 5 a cikin ɗaki da zafin jiki don cire ɗakin ɗakin karatu.
- Ƙara 5 μL na 200 mM Tris-HCl (pH 7.0) zuwa Den Tube don kawar da amsa.
- Vortex da Den Tube kuma juya shi a takaice.
- Ƙara 985 μL na prechilled HT1 zuwa Den Tube.
- Vortex da Den Tube kuma juya shi a takaice. Ana iya adana bututun a +4 ° C har sai an yi amfani da shi (a rana guda).
- Canja wurin 205 μL daga Den Tube zuwa Seq Tube.
- Ƙara 1095 μL na prechilled HT1 zuwa Seq Tube.
- Juya Seq Tube don haɗa reagents kuma juya shi ƙasa a taƙaice. Matsakaicin nauyin kaya na ƙarshe shine 1.3 ml.
- Nan da nan ci gaba zuwa 2.5 Yi Olink® gudanar da jerin gwano.
Yi Olink® gudu-gudu
A yayin wannan matakin, an ɗora kwandon buffer, tantanin da ke gudana da kuma katun da aka shirya wanda ke ɗauke da Laburaren Olink a cikin NextSeq 550, kuma ana fara aiwatar da tsarin ta amfani da girke-girke na al'ada na Olink.
Shirya benci
- Seq Tube (tare da shirye don loda ɗakin karatu), wanda aka shirya a mataki na baya
- 1x NextSeq ™ 500/550 Babban Fitarwa Reagent Cartridge v2, wanda aka shirya a mataki na baya
- 1x NextSeq ™ 500/550 Babban Fitowar Fitowa Mai Ruwa v2.5, wanda aka shirya a mataki na baya
- 1x NextSeq™ 500/550 Buffer Cartridge v2 (75 hawan keke), a dakin da zafin jiki
Saita sigogin gudu na jeri
A yayin wannan matakin, ana zaɓar sigogin gudanar da jeri akan NextSeq™ 550.
- A allon Gida na NextSeq™ 550, zaɓi Gwaji.
- A kan Zaɓan allo na tantancewa, zaɓi Jeri.
- A cikin Run Setup page, zaɓi Manual run yanayin sa'an nan gaba.
- Saita sigogin gudu kamar haka:
- A cikin filin Run Sunan, shigar da ID ɗin gwaji na musamman.
- A cikin filin ID na Library, shigar da ID na Laburare da kuke aiki (na zaɓi).
- A cikin filin 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
- MUHIMMI: Yana da mahimmanci cewa an saita karatun 1 zuwa 24, in ba haka ba duk gudu zai gaza.
- Ajiye akwatin rajistan abubuwan da ba a zaɓa ba.
- Saita wurin babban fayil ɗin fitarwa don ɗanyen bayanan gudu na yanzu. Zaɓi Bincike don canza wurin babban fayil ɗin fitarwa.
- Kada a kafa Sampda Sheet.
- Zaɓi Kayayyakin Cire don wannan gudu.
- Zaɓi Na Gaba.
Load ɗin tantanin halitta zuwa NextSeq™ 550
- Cire tantanin halitta mai gudana da aka yi amfani da shi daga gudu na baya.
- Sanya sabon tantanin halitta wanda aka shirya akan stage.
- Zaɓi Load. Ana rufe ƙofar ta atomatik.
- Lokacin da ID ɗin tantanin halitta ya bayyana akan allon kuma ana duba firikwensin a kore, zaɓi Na gaba.
Cire akwati na reagents
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 550 Jagoran Tsarin.
- Bude kofar dakin ajiyar kaya, cire kwandon da aka kashe na reagents daga žasashen, sa'annan a zubar da abun cikin daidai da ma'auni.
- Zamar da kwandon mara komai na reagent baya cikin ɓangarorin ƙarami. Dannawa mai ji yana nuna cewa an sanya akwati daidai.
Load buffer cartridge
- Cire harsashin buffer ɗin da aka yi amfani da shi daga ɗakin ajiya na sama kuma jefar da abun ciki daidai da ma'auni masu dacewa.
- Zamar da sabon kwandon buffer zuwa cikin babban ɗakin ajiya na sama. Dannawa mai ji yana nuna cewa an sanya harsashi daidai. Tabbatar cewa ID na harsashi na buffer ya bayyana akan allon kuma an duba firikwensin a kore.
- Rufe qofar rumbun adana bayanai kuma zaɓi Na gaba.
Load reagent harsashi
- Bude kofar sashin reagent, cire harsashin reagent da aka yi amfani da shi, sannan a zubar da abubuwan da ba a yi amfani da su ba daidai da ma'auni masu dacewa. Tafki a matsayi na 6 ana iya cirewa don sauƙaƙe zubar da lafiya.
- Soka hatimin tafki #10 mai lakabin "Load Library Here" tare da tukwici mai tsabta 1 ml.
- Load da 1.3 ml na Olink Library daga Seq Tube a cikin tafki #10 mai lakabin "Load Library Here".
- Zamar da sabon reagent cartridge a cikin reagents kuma rufe ƙofar reagent.
- Zaɓi Load kuma jira ~ 30 seconds har sai ID na harsashi na reagent ya bayyana akan allon kuma ana duba firikwensin a kore.
- Daga jerin abubuwan da aka saukar da girke-girke, zaɓi zaɓin girke-girke [Custom] "Olink_NSQ550_HighOutput_V1".
- MUHIMMI: Tabbatar cewa an shigar da girke-girke na al'ada a baya a cikin kayan aiki. Koma zuwa 2.2 Shigar Olink® girke-girke na al'ada.
- Zaɓi Na Gaba.
- Tabbatar da sigogin gudu da aka nuna akan Review allo. Don shirya kowane sigogi, danna Baya don komawa zuwa allon Saita Run.
- Zaɓi Na Gaba. Gudun yana farawa bayan duban riga-kafi ta atomatik. Lokacin gudanar da jeri yana kusan 7h30 min.
- MUHIMMI: Tabbatar cewa gudu yana farawa lokacin da aka kammala rajistan farko ta atomatik (~ mintuna 5).
- NOTEGa kowane gazawar rajistan da aka riga aka yi, koma zuwa umarnin masana'anta.
- NOTE: Yi hankali don kada ku shiga ciki ko in ba haka ba ku dame NextSeq™ 550 yayin gudanar da jerin abubuwan. Kayan aiki yana kula da rawar jiki.
- Tsaftace wurin aiki.
- NOTE: Lokacin da aka gama gudanar da jerin abubuwan, software ɗin tana ƙaddamar da wankin bayan-gudu ta atomatik ta amfani da maganin wankin da aka bayar a cikin kwandon buffer da NaOCl da aka bayar a cikin reagent harsashi. Wannan wankin yana ɗaukar kusan mintuna 90. Maɓallin Gida yana aiki da zarar an gama wankewa. Za a iya barin harsashin da aka yi amfani da su da tantanin halitta masu gudana a wurin har sai na gaba.
Kula da ci gaban gudu
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.
Tarihin bita
| Sigar | Kwanan wata | Bayani |
| 1.2 | 2022-11-01 | 1.2 Maye gurbin Olink® Mydata tare da Olink® NPX Explore.
2.4 Ƙara 0.2 N zuwa Kafin farawa. |
| 1.1 | 2021-12-13 | Canje-canjen Edita |
| 1.0 | 2021-12-01 | Sabo |
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
1192, v1.2, 2022-11-01
Takardu / Albarkatu
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Olink NextSeq 550 Bincika Jeri [pdf] Manual mai amfani Na gaba Seq 550 Bincika Jeri, Na gaba Seq 550, Bincika Jeri, Jeri |
