Qalabka tooska ah ee RT-qPCR
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 ah oo baakadeysan oo RT-qPCR ah iyo ensaym hufan oo sarreeyaisku dar ah (Taq, M-MLV, RI), taasoo ka dhigaysa mid ku habboon abuurista xirmooyinka dambe iyo nidaamkahagaajin, gudaha xaqiijinta fudud.
Tilmaamaha
500T, 50,000T
Qaybaha kitka
| Qalabka ku jira (Nidaamka falcelinta 20 μL) | IM-05111-01 | IM-05112-01 |
| 500 T | 50,000 T | |
| Isku-dhafka Enzyme Tooska ah | 500 μL×1 | 50 ml×1 |
| 2× Direct RT-qPCR Buffer | 1 ml×5 | 500 ml×1 |
| Tilmaanta | 1 | 1 |
Tallaabooyinka Hawlgelinta
Kahor diyaarinta reagent, dhammaan reage-yaasha ku jira xirmada waa in lagu dhalaaliyaa heerkulka qolka oo si tartiib ah loo qaso.
A: Diyaari of template iyo reajin
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), aasaasiyaal gaar ah (10 μM) iyo isticmaalka kale ee quseeya, qalab.
2. Dhig 2× Direct RT-qPCR Buffer, RNase-free ddH2O iyo 20× ROX Tixraaca Midabka (haddii ay tahay's loo baahan yahay) sanduuqa baraf leh, samaynta kuwan si dabiici ah u dhalaali, oo si tartiib ah isku qasan.
B: Diyaari of RT-qPCR nidaamka
Miis 1 : Diyaari of RT-qPCR snidaamka
| Qayb | Mugga | U fiirsashada kama dambaysta ah |
| 2× Direct RT-qPCR Buffer | 10 μl | 1× |
| Isku-dhafka Enzyme Tooska ah | 1μL | |
| Hordhac Hore (10 μM) | 0.8 μl | 50-900nM |
| Dib u rogaal celiska (10 μM) | 0.8 μl | 50-900nM |
| Baaritaan (4 μM) | 1μL | 200nM |
| Template (RNA ama Lysate) | X μL | |
| 20× ROX Tixraaca Midabka | - | 1* |
| RNase-FreeddH2O | (6.4-X) μL | |
| Wadarta mugga | 20μL |
C: Deji ah RT-qPCR falcelin barnaamijka
Miis 2 : RT-qPCR falcelinon barnaamijka dejinta (waayo tusaale: laba-tallaabo habka)
| Talaabada | Heerkulka | Waqtiga | Wareegtada | Nuxurka | |
| 1 | 50℃ | 15 daqiiqo | 1* | 1 | Dib u qorista |
| 2 | 95℃ | 1 daq | 1 | Denaturation ka hor | |
| 3 | 95℃ | 10 ilbiriqsi | 40-45 | Denaturation | |
| 60℃ | 30 ilbiriqsi 2* | Anneal/Kordhinta | |||
Kaydinta iyo nolosha shelf
-Qalabka waa in lagu kaydiyaa -20± 5℃.Muddada ansaxnimadu waa 12 bilood.
-Ka fogow wareegyada qaboojinta ee soo noqnoqda (<5 wareegyada).
