Olink nyochaa usoro site na iji NextSeq 550

Okwu mmalite

Ezubere iji
Olink® Explore bụ ikpo okwu immunoassay multiplex maka nchọpụta biomarker protein mmadụ. Ezubere ngwaahịa a maka iji nyocha naanị, ọ bụghị maka ojiji na usoro nyocha. Ọrụ ụlọ nyocha ga-abụ naanị ndị ọrụ ụlọ nyocha a zụrụ azụ. Ọ bụ naanị ndị ọrụ a zụrụ azụ ga-arụ nhazi data. Ihe nchoputa a bu n'obi ka ndị nyocha jiri ya na nchọta ụlọọgwụ ma ọ bụ ụlọ nyocha ndị ọzọ.

Banyere akwụkwọ ntuziaka a
Akwụkwọ ntuziaka onye ọrụ a na-enye ntụzịaka achọrọ ka usoro Olink® Explore Libraries na Illumina® NextSeq™ 550. Ekwesịrị ịgbaso ntuziaka ahụ nke ọma na nke doro anya. Ngbanwe ọ bụla n'ime usoro nyocha ụlọ nyocha nwere ike ibute data adịghị mma. Tupu ịmalite usoro nyocha ụlọ nyocha, lelee Olink® Explore Overview Akwụkwọ ntuziaka onye ọrụ maka mmeghe na ikpo okwu, gụnyere ozi gbasara reagents, akụrụngwa na akwụkwọ achọrọ, ihe gafereview nke usoro ọrụ, yana nke ọma dị ka ụkpụrụ ụlọ nyocha. Maka ntuziaka maka otu esi agba ọsọ Olink® Explore Reagent Kits, rụtụ aka na ntuziaka onye ọrụ Olink® Explore dị. Maka nhazi data na nyocha nke nsonaazụ usoro Olink® Explore, rụtụ aka na ntuziaka onye ọrụ Olink MyData Cloud. Ighalaahia na nwebiisinka niile dị n'ime ihe a bụ ihe onwunwe nke Olink® Proteomics AB, belụsọ ma ekwuru ya ọzọ.

Nkwado ndị teknuzu
Maka nkwado teknụzụ, kpọtụrụ Olink Proteomics na support@olink.com.

Ntuziaka ụlọ nyocha

Isiakwụkwọ a na-enye ntụzịaka maka otu esi ahazi ọba akwụkwọ Olink na NextSeq™ 550 site na iji NextSeq™ 500/550 High Output Kit v2.5 (75 Cycles). Usoro eji eme usoro bụ mmegharị nke Illumina® ọkọlọtọ NGS workflow maka Illumina® NextSeq ™ 550. Tupu ịga n'ihu na usoro, jide n'aka na enyochala ogo nke ọbá akwụkwọ a sachara. Rụtụ aka na akwụkwọ ntuziaka onye ọrụ Olink Explore dị maka ntuziaka gbasara njikwa mma.

Hazie usoro ịgba ọsọ
Enwere ike ịdezi otu ọba akwụkwọ Olink site na NextSeq™ 550 High Output flow cell na kwa ọsọ. Ọnụ ọgụgụ nke mkpụrụ ndụ na-agba ọsọ dị elu na-agba ọsọ chọrọ ka usoro ihe dị iche iche Olink Explore Reagent Kits ka akọwara na Tebụl 1. Ọ bụrụ na achọrọ ihe karịrị otu ọsọ, megharịa ntụziaka ndị akọwapụtara na ntuziaka a.

Tebụl 1. Usoro nhazi usoro ịgba ọsọ:

Ngwa Olink® Explore Reagent	Ọnụọgụ nke ọba akwụkwọ Olink	Ọnụọgụ cell (s) na ọsọ(s) eruba
Olink® Explore 384 Reagent Kit	1	1
Olink® Explore 4 x 384 Reagent Kit	4	4
Olink® Explore 1536 Reagent Kit	4	4
Ngwa Olink® nyocha Mgbasawanye Reagent Kit	4	4
Olink® Explore 3072 Reagent Kit	8	8

Wụnye uzommeputa omenala Olink
N'oge usoro a, a na-etinye usoro nhazi omenala Olink® na NextSeq™ 550. Nzọụkwụ a kwesịrị ka eme otu ugboro, tupu emee usoro nhazi Olink maka oge mbụ.
IHE: Ntụziaka omenala Olink ga-arụ ọrụ naanị na NextSeq ™ 500/550 Ngwa mmepụta dị elu yana NextSeq ™ Control Software 4.0

1. Wepu ma tinye usoro nhazi omenala Olink Olink_NSQ550_HighOutput_V1 n'ime folda na-esote nke NextSeq™ 550 ngwá: C:\Program Files\IluminaNextSeq Control SoftwareRecipeCustomHigh\.
2. N'okpuru ahaziri Sistemu> Jikwaa akụrụngwa, mee ka Ezi ntụziaka Omenala. Ọ bụrụ na ahọpụtaghị ya, nhọrọ nhazi omenala agaghị apụta n'oge nhazi ọsọ.

IHE: Na ụdị sọftụwia NCS 4.0, nhọrọ ịhọrọ usoro nhazi omenala ga-eme naanị mgbe ebuchara katrij reagent, ọ bụghị na ibe ntọala mbụ.
IHE: A ghaghị ịtọ ọsọ ahụ n'ụdị akwụkwọ ntuziaka iji kwe ka ntụzịaka omenala.

Kwadebe reagents usoro

N'oge usoro a, a na-agbaze katrij reagent nke nwere nchịkọta na usoro reagents ma kwadebere cell eruba.

Kwadebe katrij reagent
Ịdọ aka ná ntị: Katrij reagent nwere kemịkalụ nwere ike ịdị ize ndụ. Yiri akụrụngwa nchebe zuru oke ma tụfuo reagents eji eme ihe dịka ụkpụrụ dabara adaba. Maka ozi ndị ọzọ, rụtụ aka na Ntuziaka Sistemụ Illumina NextSeq 550 (akwụkwọ #15069765).

Kwadebe oche

• 1x NextSeq ™ 500/550 Reagent Cartridge v2 (usoro okirikiri 75).

Ntuziaka

1. Tinye katriji reagent oyi kpọnwụrụ ọkara-imerime n'ime ụlọ mmiri dị ọkụ wee hapụ ya ka ọ gbazee maka elekere 1. Gbaa mbọ hụ na agbazere katrij niile nke reagent kpamkpam.
   IHE: Maka ịdị mma, kpochaa katriji ahụ ụbọchị gara aga ma chekwaa ya n'abalị na 4 Celsius C. Na okpomọkụ a, reagents kwụsiri ike ruo otu izu.
2. Jiri akwa mpempe akwụkwọ kpoo ntọala katrij ahụ nke ọma wee hichapụ ihe mkpuchi foil ahụ site na anụ ahụ na-enweghị lint ma ọ bụrụ na ọ dị mkpa.
3. Tụgharịa katrij ugboro iri ka mix nke ọma reagents thawed n'ime.
4. Kpatụ katriji ahụ nwayọ n'oche iji wepụ ikuku ikuku. Chekwaa katrij na ụlọ okpomọkụ ma ọ bụrụ na a ga-eji ya n'ime awa 4.

Kwadebe cell eruba

Kwadebe oche

• 1x NextSeq ™ 500/550 Igwe nrụpụta dị elu v2.5.

Ntuziaka

1.  Weta cell refrigerated n'ime ụlọ okpomọkụ maka nkeji 30.
2.  Yiri uwe aka na-enweghị ntụ ntụ ọhụrụ (iji zere imerụ elu iko nke cell na-eru eru).
3.  Mgbe ị dị njikere ibunye cell eruba n'ime ngwá ọrụ ahụ, wepụ mkpụrụ ndụ na-asọpụta na ngwugwu yana clamshell rọba.
4.  Nyochaa cell eruba. Ọ bụrụ na a na-ahụ akụkụ ma ọ bụ uzuzu n'elu ihe ọ bụla n'ime iko ahụ, hichaa elu ọdabara na mmanya isopropyl na-enweghị lint ma jiri anụ ụlọ nyocha dị ala kpoo ya.

Kwadebe Ọbá akwụkwọ Olink maka usoro

N'oge usoro a, a na-akwado dilutions NaOH na Tris-HCl, yana ọbá akwụkwọ Olink a na-achịkwa nke dị ọcha na nke dị mma na-agbaze ma wepụ ya n'usoro usoro.

Kwadebe NaOH dilution
A na-eji mgbaze NaOH iji denature ụlọ akwụkwọ.

Kwadebe oche

• 1 N NaOH ngwaahịa
• Mmiri MilliQ
• 1 x Microcentrifuge tube (1.5 ml)
• Pipette akwụkwọ ntuziaka (10-100 μL)
• Filter ndụmọdụ pipette

Tupu ịmalite
Kaa akara microcentrifuge tube "NaOH".

Ntuziaka

1. Kwadebe 0.2 N NaOH dilution na NaOH Tube dị ka Tebụl 2 si dị.
2. Vortex na NaOH Tube nke ọma wee gbadaa. Jiri n'ime awa iri na abụọ.

Isiokwu 2. 0.2 N NaOH dilution

Reagent	Olu (μL)
Mmiri MilliQ	80
1 N NaOH ngwaahịa	20

2.4.2 Kwadebe Tris-HCl dilution
A na-eji mgbaze Tris-HCl mee ka ọ ghara imebi ọba akwụkwọ denaturated.

Kwadebe oche

• 1 M Tris-HCl pH 7.0 ngwaahịa (Trizma® hydrochloride ngwọta)
• Mmiri MilliQ
• 1 x Microcentrifuge tube (1.5 ml)
• Pipette akwụkwọ ntuziaka (10-100 μL)
• Filter ndụmọdụ pipette

Tupu ịmalite

• Kaa akara microcentrifuge tube "Tris-HCl"

Ntuziaka

1. Kwadebe 200mM Tris-HCl dilution na Tris-HCl Tube dị ka Tebụl 3 si dị.
2. Vortex Tris-HCl Tube nke ọma wee tụgharịa ya.

Isiokwu 3. 200 mM Tris-HCl dilution:

Reagent	Olu (μL)
Mmiri MilliQ	80
1M Tris-HCl pH 7.0 ngwaahịa (Trizma® hydrochloride ngwọta)	20

Mebie ọba akwụkwọ Olink®
N'oge usoro a, ọbá akwụkwọ Olink a na-achịkwa nke dị ọcha na nke dị mma na-agbaze 1:33.

Kwadebe oche

• Lib Tube, akwadoro dịka akwụkwọ ntuziaka onye ọrụ Olink Explore siri dị
• Mmiri MilliQ
• 1 x Microcentrifuge tube (1.5 ml)
• Pipettes akwụkwọ ntuziaka (0.5-10 na 100-1000 μL)
• Filter ndụmọdụ pipette

Tupu ịmalite

• Mepee tube Lib ma ọ bụrụ oyi.
• Kaa akara tube microcentrifuge ọhụrụ: "Dil".

Ntuziaka

1. Tinye 96 μL nke mmiri MilliQ na Dil Tube.
2. Vortex na Lib Tube ma tụgharịa ya obere oge.
3. Nyefee 3 μL site na Lib Tube na Dil Tube.
4. Vortex na Dil Tube ma tụgharịa ya obere oge

Denature na itughari Ọbá akwụkwọ Olink na njedebe ikpeazụ
N'oge usoro a, a na-ewepụ ọbá akwụkwọ Olink nke agbazere ma tinyekwuo ya na ntinye ntinye ikpeazụ.

Kwadebe oche

• Dil Tube, akwadoro na nzọụkwụ gara aga
• 0.2 N NaOH dilution, akwadoro ọhụrụ na nzọụkwụ gara aga
• 200 mM Tris-HCl (pH 7.0) dilution, kwadebere na nzọụkwụ gara aga
• Ihe nchekwa ngwakọ 1 (HT1) gụnyere na igbe Ngwa Ngwa NextSeq 2
• 2 x Microcentrifuge tubes (1.5 ml na 2 ml)
• Pipettes akwụkwọ ntuziaka (0.5-10 na 100-1000 μL)
• Filter ndụmọdụ pipette

Tupu ịmalite

• Kpoo ihe nchekwa HT1 oyi kpọnwụrụ n'ime ụlọ okpomọkụ. Chekwaa na +4 Celsius C ruo mgbe ejiri ya.
• Kanye akara tube microcentrifuge ọhụrụ 1.5 ml: “Den” (maka ọba akwụkwọ denaturated).
• Kaa akara tube microcentrifuge 2 ml ọhụrụ ahụ: “Seq” (maka onye dị njikere ibu ọba akwụkwọ).

Ntuziaka

1. Nyefee 5 μL site na Dil Tube na Den Tube.
2. Tinye 5 μL nke 0.2 N NaOH n'ime oghere oghere.
3. Vortex na Den Tube ma tụgharịa ya obere oge.
4. Tinye ọwa mmiri ahụ maka nkeji 5 n'ime ụlọ okpomọkụ ka ọ ghara imebi ụlọ akwụkwọ ahụ.
5. Tinye 5 μL nke 200 mM Tris-HCl (pH 7.0) na Den Tube iji wepụ mmeghachi omume ahụ.
6. Vortex na Den Tube ma tụgharịa ya obere oge.
7. Tinye 985 μL nke HT1 akpọchiri akpọchi na Ọwa Ọgba.
8. Vortex na Den Tube ma tụgharịa ya obere oge. Enwere ike ịchekwa tube ahụ na +4 Celsius C ruo mgbe ejiri ya (otu ụbọchị).
9. Nyefee 205 μL site na Den Tube na Seq Tube.
10. Tinye 1095 μL nke HT1 prechilled na Seq Tube.
11. Tụgharịa Seq Tube ka ịgwakọta reagents wee tụgharịa ya obere oge. Olu ikpeazụ bụ 1.3 ml.
12. Ngwa ngwa gaa n'ihu na 2.5 Mee usoro usoro Olink®.

Mee usoro usoro Olink®

N'ime usoro a, a na-ebunye katrij buffer, cell flow na katrij reagent a kwadebere nke nwere Ọbá akwụkwọ Olink n'ime NextSeq 550, a na-amalite usoro nhazi site na iji nhazi omenala Olink.

Kwadebe oche

• Seq Tube (nwere njikere ibu ọba akwụkwọ), akwadoro na nzọụkwụ gara aga
• 1x NextSeq ™ 500/550 Reagent Cartridge v2, akwadoro na nzọụkwụ gara aga
• 1x NextSeq ™ 500/550 Igwe nrụpụta dị elu v2.5, akwadoro na nzọụkwụ gara aga
• 1x NextSeq™ 500/550 Buffer Cartridge v2 (usoro okirikiri 75), na okpomọkụ ụlọ

Tọọ usoro ọsọ ọsọ
N'oge usoro a, a na-ahọrọ usoro ọsọ ọsọ na NextSeq™ 550.

1. Na NextSeq™ 550 Ihuenyo mbu, họrọ nnwale.
2. Na ihuenyo Họrọ Assay, họrọ Usoro.
3. Na Run Mbido ibe, họrọ Manual run mode wee na-esote.
4. Tọọ paramita ọsọ ọsọ dị ka ndị a:
   • Na ọsọ Aha ubi, tinye a pụrụ iche nnwale NJ.
   • N'ime oghere ID Library, tinye ID nke Library ị na-agba (nhọrọ).
   • Na Gụọ Ụdị ubi, họrọ Single-agụ nhọrọ.
   • Tinye ọnụọgụ okirikiri dịka ndị a:
     • Gụọ 1:24
     • Ndekọ 1: 0
     • Ndekọ 2: 0
     • Gụọ 2:0
   • Debe igbe nrịbama maka primers omenala na-ahọrọghị.
   • Tọọ ebe nchekwa mmepụta maka data raw na-agba ọsọ ugbu a. Họrọ Chọgharịa ka ịgbanwee ọnọdụ nchekwa mmepụta.
5. Ewelitela Sample Mpempe akwụkwọ.
6. Họrọ Hichapụ ihe eji eme ihe maka ọsọ a.
7. Họrọ Ọzọ.

Budata cell eruba n'ime NextSeq™ 550

1. Wepu cell mgbaba ejiri n'ọsọ gara aga.
2. Debe cell eruba akwadoro ọhụrụ na stage.
3. Họrọ Ibu. A na-emechi ụzọ na-akpaghị aka.
4. Mgbe NJ cell eruba pụtara na ihuenyo wee lelee sensọ na akwụkwọ ndụ akwụkwọ ndụ, họrọ Ọzọ

Gbanyụọ akpa reagent
Ịdọ aka ná ntị: Ntọala reagents a nwere kemịkalụ nwere ike ịdị ize ndụ. Yiri akụrụngwa nchebe zuru oke ma tụfuo reagents eji eme ihe dịka ụkpụrụ dabara adaba. Maka ozi ndị ọzọ, rụtụ aka na Ntuziaka Sistemụ Illumina NextSeq 550.

1. Mepee ọnụ ụzọ nke ebe nchekwa, wepụ akpa reagents n'ime obere ụlọ, ma tụfuo ọdịnaya dị ka ụkpụrụ dabara adaba.
2. Weghachite akpa reagenti efu azụ n'ime ebe nchekwa ala. Ọpịpị a na-anụ na-egosi na etinyere akpa ahụ nke ọma.

Buffer katrij

1. Wepu katrij ihe nchekwa eji eme ihe site na ngalaba nchekwa elu wee tụfuo ọdịnaya dịka ụkpụrụ dabara adaba.
2. Manye katrij ihe nchekwa ọhụrụ n'ime ebe nchekwa nke elu. Ọpịpị a na-anụ na-egosi na etinyere katrijị ahụ nke ọma. Gbaa mbọ hụ na NJ katrij nke buffer na-egosi na ihuenyo yana na a na-enyocha sensọ na akwụkwọ ndụ akwụkwọ ndụ.
3. Mechie ọnụ ụzọ nke ebe nchekwa wee họrọ Ọzọ.

Ibu ibu reagent katrij

1. Mepee ọnụ ụzọ nke reagent, wepụ katrij reagent ejiri mee ihe, ma tụfuo ọdịnaya ejighi ya dịka ụkpụrụ dabara adaba. Ebe nchekwa nchekwa dị n'ọnọdụ 6 bụ nke mbughari iji kwado mkpofu nchekwa.
2. Gbuo akara nke ọdọ mmiri #10 nke akpọrọ dị ka "Load Library Here" na n'ọnụ pipette 1 ml dị ọcha.
3. Bunye 1.3 ml Ọbá akwụkwọ Olink site na Seq Tube n'ime mmiri mmiri #10 akpọrọ "Load Library Here".
4. Wepụ katrij reagent ọhụrụ n'ime ụlọ reagent ma mechie ọnụ ụzọ nke reagent.
5. Họrọ Ibu ma chere maka ~ 30 sekọnd ruo mgbe ID katrij reagent pụtara na ihuenyo wee lelee sensọ na akwụkwọ ndụ akwụkwọ ndụ.
6. Site na ndetu ndetu usoro nri, họrọ nhọrọ uzommeputa [Custom] “Olink_NSQ550_HighOutput_V1.
7. Họrọ Ọzọ.

Malite usoro ịgba ọsọ

1. Kwenye usoro ịgba ọsọ egosiri na Review ihuenyo. Iji dezie paramita ọ bụla, pịa Azụ ka ịlaghachi na ihuenyo ọsọ ntọala.
2. Họrọ Ọzọ. Ọsọ na-amalite mgbe nlele mbụ emechara ya. Oge ịgba ọsọ usoro bụ ihe dịka 7h30 min.
3. Hichaa ebe ọrụ.

IHE: Mgbe emechara usoro ihe omume ahụ, ngwanro ahụ na-amalite ịsacha akpaaka na-akpaghị aka site na iji ngwọta ịsacha enyere na katrij buffer yana NaOCl enyere na katrij reagent. Ịsacha a na-ewe ihe dịka nkeji iri itoolu. Bọtịnụ ụlọ na-arụ ọrụ ozugbo ịsachara ya. Enwere ike ịhapụ katrij ndị ejiri mee ihe na cell eruba n'ebe ahụ ruo oge ọzọ.

Nyochaa ọganihu ịgba ọsọ
Olink na-eji NGS dị ka agụpụta iji tụọ ọnụọgụ nke usoro a ma ama iji tụọ mkpokọta protein enyere na s.amples (n'ihe metụtara ndị ọzọ samples). A na-ekpebi ogo data sitere na ọsọ usoro nyocha nke ọ bụla site na paramita QC pụrụ iche na teknụzụ Olink. N'ihi ya, metrik njikwa ogo ọkọlọtọ ejiri na NGS, dị ka Q-score, adịchaghị mkpa.

Akụkọ ngbanwe

Ụdị	Ụbọchị	Nkọwa
1.1	2021-12-13	Mgbanwe ederede
1.0	2021-12-01	Ọhụrụ

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Olink Proteomics AB nwere akara ụghalaahịa a: Olink®.
Ekpuchiri ngwaahịa a n'ọtụtụ patent na ngwa patent dị na https://www.olink.com/patents/.
© Copyright 2021 Olink Proteomics AB. Akara ụghalaahia ndị ọzọ bụ ihe onwunwe nke ndị nwe ha. Olink Proteomics, Dag Hammarskjölds na 52B , SE-752 37 Uppsala, Sweden
1192, v1.1, 2021-12-13

Akwụkwọ / akụrụngwa

Olink nyochaa usoro site na iji NextSeq 550 [pdf] Akwụkwọ ntuziaka onye ọrụ Nyochaa usoro site na iji NextSeq 550, nyochaa usoro, na-eji NextSeq 550, NextSeq 550

Ntụaka

• Akwụkwọ ntuziaka onye ọrụ