Tooska RT-qPCR Kit III
Sharaxaada Qalabka:
◮Qalabka tooska ah ee RT-qPCR, iyadoo la adeegsanayo Foreasy Reverse Transcriptase iyo Foreasy HS Taq DNA Polymerase ee uu soo saaray Foregene, oo leh falcelin gaar ah, ayaa ka dhigaysa xirmadan iska caabin xooggan iyo waafaqid, waxayna si toos ah u tijaabin kartaa falcelinta adeegsiga nidaamka Foregene Lysis qaab qaab ahaan.Qalabkani waxa uu dalban karaa xirmada ogaanshaha aashitada nucleic ee COVID-19 iyo horumarinta xirmada ogaanshaha asiidhka kale ee bakteeriyada.
Sharaxaada
Qalabka tooska ah ee RT-qPCR wuxuu bixiyaa baakad gooni gooni ah u xirxiran ee RT-qPCR (oo ay ku jiraan Foreasy HS Taq DNA polymerase isku dhafan) iyo tayo sare lehQoraal-qorista Dib-u-noqoshada Saadaasha (MMLV),taasoo ka dhigaysa mid ku habboon abuurista xirmooyinka dambe-dhamaadka iyo hagaajinta nidaamka ， gudahooda xaqiijinta fudud.
Qalabka tooska ah ee RT-qPCR, iyadoo la adeegsanayo Foreasy Reverse Transcriptase iyo Foreasy HS Taq DNA Polymerase ee uu soo saaray Foregene, oo leh falcelin gaar ah, ayaa ka dhigaysa xirmadan iska caabin xooggan iyo waafaqid, waxayna si toos ah u tijaabin kartaa falcelinta adeegsiga nidaamka Foregene Lysis qaab qaab ahaan.Qalabkani waxa uu dalban karaa xirmada ogaanshaha aashitada nucleic ee COVID-19 iyo horumarinta xirmada ogaanshaha asiidhka kale ee bakteeriyada.
Qalabkani wuxuu si toos ah u noqon karaa qaybta alaabta IVD, iyadoo la isticmaalayo si fudud oo degdeg ah.Waxay kaliya u baahan tahay xaqiijin sahlan, iyada oo aan dib loo horumarin.
Tilmaamaha
500T, 50,000T
Qaybaha kitka
| Qalabka ku jira (Nidaamka falcelinta 20 μL) | IM-05111-03 | IM-05112-03 |
| 500 T | 50,000 T | |
| Hordhac REverse Transcriptase(200 U/μL) | 50 μL×1 | 1 ml×5 |
| 2× QPCR toos ahMix-Taqman | 1 ml ×5 | 500 ml×1 |
| Tilmaanta | 1 | 1 |
Tallaabooyinka Hawlgelinta
A: Diyaari template iyo reagent
1. Diyaari template RNA diyaarsan (Waxaa lagu talinayaa in la isticmaalo Foregene Total RNA Go'doomin Kit taxane xirmo si loo soo saaro oo loo nadiifiyo template RNA) ama alaabta dildilaaca muunad (Waxaa lagu talinayaa in la isticmaalo Foregene Lysis nidaamka), aasaasiga ah (10 μM) gaar ah (10 μM) iyo isticmaalka kale ee quseeya, qalab.
2. Ku rid 2× Direct RT-qPCR Buffer, RNase-free ddH2O iyo 20× ROX Reference Dye(haddii loo baahdo) sanduuqa baraf leh, adigoo kuwan u dhalaalinaya si dabiici ah, oo si tartiib ah isku qasan.
B: Diyaarinta nidaamka RT-qPCR
Hel kala badh mugga falcelinta celiska ee habka falcelinta (tusaale, haddii mugga nidaamku yahay 20 μL, waxa uu u baahan yahay in uu helo 10 μL 2× Direct RT-qPCR Buffer). Waayoqadarka u dhiganta ee enzyme,waxaanu soo jeedinaynaaM-MLV: 10-30 U/20 μL nidaamka falcelinta,Taqpolymerase DNA：1-2 U/20 μL habka falcelintatani waa soo jeedinteena, waa inaad tijaabisaa oo aad hagaajisaa), ku dar template RNA, aasaasiyaal gaar ah iyo baarayaal, oo ku dar RNase-free ddH2O ilaa 20 μL.Diyaarinta gaarka ah ee nidaamka falcelinta RT-qPCR waxaa loo tixraaci karaa shaxda 1 ee soo socota.
Shaxda 1: Diyaarinta nidaamka RT-qPCR
| Qayb | Mugga | U fiirsashada kama dambaysta ah |
| 2× Direct RT-qPCR Buffer | 10 μl | 1× |
| Qoraal-buuxinta hore (MMLV) | 10-30 U | |
| Horudhac HS Taq DNA Polymerase | 1-2 U | |
| Hordhac Hore (10 μM) | 0.8 μl | 50-900nM |
| Dib u rogaal celiska (10 μM) | 0.8 μl | 50-900nM |
| Baaritaan (4 μM) | 1 μL | 200nM |
| Template (RNA ama Lysate) | X μL | |
| 20× ROX Tixraaca Midabka | - | 1* |
| RNase-FreeddH2O | (6.4-X) μL | |
| Wadarta mugga | 20 μl |
Fiiro gaar ah: Horudhac Primer iyo Reverse Primer waa aasaasayaal gaar ah oo hiddo-wadaha bartilmaameedka ah.Nidaamka qPCR waa la hagaajin karaa iyadoo loo eegayo tijaabooyinka la taaban karo iyo moodooyinka PCR.Waxaan ku talinaynaa 400nM fiirsashada kama dambaysta ah ee aasaasayaasha intooda badan.Hagaajin mugga aasaasiyaal gaar ah iyo baarayaal iyadoo loo eegayo diiradda la diyaariyey iyo diiradda ugu dambeysa ee lagu taliyey, fadlan.
1*: Dooro fiirsashada kama dambaysta ah ee ROX Reference Dye iyadoo loo eegayo qalabyada PCR ee kala duwan.Xoojinta dheeha tixraaca ROX ee ugu wanagsan ee qalabka PCR tirada guud ayaa lagu muujiyay shaxda soo socota:
| Qalabka PCR ee tirada leh | U fiirsashada kama dambaysta ah ee tixraaca ROX |
| ABI PRISM 7000/7300/7700/7900HT/Tallaabo Koowaad,iwm. | 1 × (tusaale ahaan, ku dar 1μl 20×ROX dheeha tixraaca ee nidaamka falcelinta 20 μL) |
| ABI 7500, 7500 Fast, Stratagene Mx3000P, Mx3005P iyo Mx4000, iwm. | 0.5× (tusaale ahaan, ku dar 0.5μl 20×ROX dheeha tixraaca ee 20 μL nidaamka falcelinta) |
| Qalabka Roche PCR, Qalabka Bio-Rad PCR, Qalabka PCR ee tirada Eppendorf, iwm. | Iyadoo aan lahayn dheeha tixraaca ROX |
Kaydinta iyo nolosha shelf
Qalabka waa in lagu kaydiyaa at -20 ± 5 ℃.Waa in isla markiiba lagu kaydiyaa -20 ℃ qaboojiyaha heerkulbeegga.Muddada ansaxnimadu waa 2 sano haddii lagu kaydiyo xaalad ku habboon.
