QuickEasyᵀᴹ Qalabka PCR ee Kit-Taqman
Sharaxaada Qalabka:
◮Nidaamka kobcinta PCR ee gaarka ah ayaa ka dhigaya 2 × QuickEasyTMReal PCR Mix-Taqman oo la jaan qaadi kara.
◮Bilawga Kulul ee Foregene HS Taq Polymerase waxa uu leeyahay hufnaan sare-u-qaadis, dareen-weyneyn sare, iyo gaar-ahaaneed sareeye.
◮2× Degdega fududTMReal PCR Mix -Taqman waxay isticmaashaa nidaam gaar ah oo ay hagaajisay Foregene si ay u wanaajiso dareenka iyo gaar ahaaneed ee ogaanshaha baaritaanka gaarka ah ee taxanaha ah, kaas oo loo isticmaali karo genotyping iyo koobiga go'aaminta kala duwanaanshaha lambarka oo loo helo natiijooyin sax ah.
◮Alaabtani waxay la socotaa dheeha tixraaca gudaha ee ROX, kaas oo loo isticmaali karo in lagu baabi'iyo asalka ishaarada iyo cilada u dhaxaysa ceelasha, taas oo ku haboon macaamiisha inay u isticmaalaan noocyada kala duwan ee qalabka PCR ee tirada.
Sharaxaada
2× QuickEasyTMReal PCR Mix-Taqman oo ay bixiso QuickEasyTMQalabka PCR ee Kit-Taqman-ka-dhabta ah waa nidaam isku-dhafan oo jiil cusub ah oo loogu talagalay fal-celinta falcelinta PCR-ga ee waqtiga-dhabta ah iyadoo la adeegsanayo baaritaanno gaar ah.Aasaaskeeda Foregene HS Taq DNA Polymerase waxay ku salaysan tahay wax ka beddelka Antibody-ka, oo lala isticmaalo Foregene nidaamka PCR ee la hagaajiyay, wuxuu leeyahay gaar gaar ah iyo hufnaan sare.Marka la barbar dhigo isku dhafka PCR ee caadiga ah, waxay leedahay awood xoojin xoog leh iyo hoos u dhac ku yimaada heerka isbarbardhigga.Waxaa loo isticmaali karaa falcelinta PCR ee cabirka fluorescence si loo yareeyo weyneynta aan gaarka ahayn loona wanaajiyo saxnaanta PCR.Waxay si weyn u wanaajin kartaa gaar ahaan badeecada iyo dareenka falcelinta.Isla mar ahaantaana, ROX waxaa loo bixiyay dheeha tixraaca gudaha ah.
The QuickEasyTMQalabka PCR ee Kit-Taqman ee waqtiga-dhabta ah waxa uu leeyahay baaxad ballaadhan oo codsiga xaddiga template ah, waxana uu heli karaa qalooca heerka wanaagsan ee gobollo tiro badan, oo si sax ah u qiyaasi kara oo ogaan kara hiddo-wadaha bartilmaameedka leh ee soo noqnoqonaya iyo kalsooni sare.
Tilmaamaha
200 oo diyaari,500 oo diyaari,1000 diyaarin,2000 diyaarin
Qaybaha kitka
| Degdega fududTMWaqtiga dhabta ah PCR Kit-Taqman(20μlnidaamka) | ||||
| Qalabka ku jira | QP-01121 | QP-01122 | QP-01123 | QP-01124 |
| 200 diyaarin | 500 diyaarin | 1000 Diyaarin | 2000 diyaarin | |
| 2×Degdeg ahFududTM PCR dhabta ah Isku qas-Taqmaan | 1 ml×2 | 1.7 ml×3 | 1.7 ml×6 | 1.7 ml×12 |
| 20×ROX Tixraaca Midabka | 200μL | 0.5 ml | 1 ml | 1 ml×2 |
| DNA-free ddH2O | 1.7 ml | 1.7 ml×2 | 10 ml | 20 ml |
| IFU | 1gabal | 1gabal | 1gabal | 1gabal |
Tilmaamaha & faa'iidooyinka
■ Nidaamka hagaajinta PCR ee gaarka ah wuxuu ka dhigayaa 2× QuickEasyTMReal PCR Mix-Taqman oo la jaan qaadi kara.
■ Hot-star Foregene HS Taq Polymerase waxay leedahay hufnaan sare-u-qaadis, dareen-weyneyn sare, iyo gaar-qaadid sare.
■ 2× Degdega fududTM Real PCR Mix -Taqman waxay isticmaashaa nidaam gaar ah oo ay hagaajisay Foregene si ay u wanaajiso dareenka iyo gaar ahaaneed ee ogaanshaha baaritaanka gaarka ah ee taxanaha ah, kaas oo loo isticmaali karo genotyping iyo koobiga go'aaminta kala duwanaanshaha lambarka oo loo helo natiijooyin sax ah.
■ Alaabadani waxay la socotaa roodhida tixraaca gudaha ee ROX, kaas oo loo isticmaali karo in lagu baabi'iyo asalka ishaarada iyo cilada u dhaxaysa ceelasha, taas oo ku haboon macaamiisha inay u isticmaalaan noocyada kala duwan ee qalabka PCR ee tirada badan.
Codsiga qalabka
■ Fluorescence quantitative PCR for template analysis analysis;
■ CKordhinta PCR ee caadiga ah;
■ Waxa loo isticmaali karaa ogaanshaha allele.
Kaydinta iyo nolosha shelf
Xirmada waa in lagu kaydiyaa -20°C gudcurka;haddii si joogto ah loo isticmaalo, waxa kale oo lagu kaydin karaa 4°C muddo gaaban (la isticmaalo 10 maalmood gudahood).
Mabaadi'da naqshadaynta aasaasiga ah ee waqtiga dhabta ah PCR
Horudhac Horudhac iyo Horudhac Hordhac ah
PCR-da-waqtiga-dhabta ah, naqshadaynta asaasiga ahi aad bay muhiim u tahay.Furayaasha waxay la xiriiraan gaarnimada iyo hufnaanta kordhinta PCR, waxaana loo qaabayn karaa iyadoo la tixraacayo mabaadi'da soo socota:
Dhererka hore: 18-30bp.
Waxa ku jira GC: 40-60%.
Qiimaha Tm: Software-ka naqshadaynta aasaasiga ah, sida Primer 5, wuxuu ku siin karaa qiimaha Tm ee asaasiga ah.Qiimayaasha Tm ee hore iyo kuwa hoose waa inay ahaadaan kuwa ugu dhow ee suurtogalka ah.Qaaciddada xisaabinta Tm waxa kale oo la isticmaali karaa: Tm = 4 °C (G + C) + 2 °C (A + T).Marka la samaynayo PCR, heerkulka ka hooseeya qiimaha Tm ee aasaasiga ah ee 5 °C ayaa guud ahaan loo doortaa heer kulka soo jiidashada (kordhinta u dhiganta heerkulka xididada ayaa kordhin kara qeexida falcelinta PCR).
Qalabka aasaasiga ah iyo PCR:
Naqshadeynta asaasiga ah ee PCR dhererka alaabada ayaa la door bidaa 100-150bp.
Naqshadeynta aasaasiga ah ee aagga qaab dhismeedka sare ee template waa in laga fogaadaa sida ugu macquulsan.
Ka fogow samaynta 2 ama in ka badan oo saldhigyo dhammaystiran oo u dhexeeya 3′ daraf ee hore-u-socodyada sare iyo kuwa hoose.
Primer 3′ terminal saldhiga kuma jiri karo 3 dheeraad ah oo isku xigta G ama C.
Furayaasha laftoodu ma yeelan karaan dhismayaal is kaaba ah, haddii kale qaab-dhismeedka timaha ayaa la samayn doonaa, oo saamaynaya kor u qaadida PCR.
ATCG waa in loo qaybiyaa sida ugu siman ee suurtogalka ah ee isku xigxiga asaasiga ah, iyo 3′ saldhigga terminal waa in laga fogaadaa sida T.
Lifaaqa 1: Xidhmada kabka qaybta gacanta tooska ah ee RT-qPCR
1.Cell Lysis Solution
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| Qaybaha kitka (24-hab lysis / ceel) | DRT-01011-A1 | DRT-01011-A2 | |
| 100 T | 500 T | ||
| QaybI | Buffer CL | 20 ml | 100 ml |
| Foregene Protease Plus II | 400 μl | 1 ml × 2 | |
| Buffer ST | 1 ml × 2 | 10 ml | |
| QaybII | Tirtiraha DNA | 400 μl | 1 ml × 2 |
2.RT Isku darka
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| Qaybaha kitka (Nidaamka falcelinta 20 μl) | DRT-01011-B1 |
| 200 T | |
| 5× Toos ah RT Mix | 800 μl |
| RNase-free ddH2O | 1.7 ml × 2 |
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| Qaybaha kitka (Nidaamka falcelinta 20 μl) | DRT-01011-C1 | DRT-01011-C2 |
| 200 T | 1000 T | |
| 2× Toos ah qPCR Mix-SYBR | 1 ml × 2 | 1.7 ml × 6 |
| 50× ROX Tixraaca Midabka | 40 μl | 200 μl |
| RNase-free ddH2O | 1.7 ml | 10 ml |
