Qalabka tooska ah ee RT-qPCR
Sharaxaada Qalabka:
◮Wixii RNA toos ah-waqtiga-dhabta ah u kordhinta ururinta suufka iyada oo aan lahayn hababka sifaynta RNA.Qalabkani waxa uu ku dhamaystirayaa wareegyada qRT-PCR hal saac gudaheed.Isku darka enzyme waa isku dhafka la hagaajiyay ee Reverse Transcriptase, Hot-Start Taq DNA polymerase, horjoogaha RNase.Reaction Buffer wuxuu ka kooban yahay dhammaan qaybaha loo baahan yahay, oo ay ku jiraan qaybaha kaydinta la hagaajiyay, Mg2+, dUTP, iyo dNTPs.
Sharaxaada
Qalabka 'ForeDirect RT-qPCR' waxaa loogu talagalay RNA si toos ah u-xoojinta-waqtiga-dhabta ah ee ururinta suufka iyada oo aan la helin hababka sifaynta RNA.Qalabkani waxa uu ku dhamaystirayaa wareegyada qRT-PCR hal saac gudaheed.Isku darka enzyme waa isku dhafka la hagaajiyay ee Reverse Transcriptase, Hot-Start Taq DNA polymerase, horjoogaha RNase.Reaction Buffer wuxuu ka kooban yahay dhammaan qaybaha loo baahan yahay, oo ay ku jiraan qaybaha kaydinta la hagaajiyay, Mg2+, dUTP, iyo dNTPs.
Tilmaamaha
100T
Qaybaha kitka
| Wakiilka sii daaya acid nucleic |
| difaaca RNA |
| 2× RT-qPCR Buffer |
| Isku-dhafka Enzyme (Taq&M-MLV) |
| Tilmaamaha |
Codsiga PCR
1. Marka tubooyinka falcelinta PCR si kooban loo xakameeyo, ku rid haanta muunada ee qalabka wax-weynaynta.
2. Hab maamuuska kulaylka
| Talaabada | Heerkulka | Waqtiga | Wareegtada | |
| 1 | Dib u qorista | 50℃ | 15 daqiiqo | 1 |
| 2 | Denaturation ka hor | 95℃ | 1 daq | 1 |
| 3 | Denaturation | 95℃ | 10 ilbiriqsi | 42 |
| 4 | Anneal/Kordhinta | 60℃ | 30 ilbiriqsi | |
Fiiro gaar ah:Digniinta fluorescent ayaa la ogaaday isla markiiba ka dib markii la kordhiyey wareeg kasta.
3. Kadib dejinta, kaydi faylka oo socodsii barnaamijka falcelinta.
Tallaabooyinka Hawlgelinta
Kahor diyaarinta reagent, dhammaan reage-yaasha ku jira xirmada waa in lagu dhalaaliyaa heerkulka qolka oo si tartiib ah loo qaso.
Diyaarinta reagent
1. Diyaari isku darka Sii deynta Nucleic Acid
| Qayb | Mugga halkii Tusaalaha ama Xakamaynta |
| Wakiilka sii daaya acid nucleic | 5μL |
| difaaca RNA | 0.5 μl |
2. Diyaari isku darka falcelinta
| Qayb | Mugga halkii Tusaalaha ama Xakamaynta |
| 2× RT-qPCR Buffer | 15 μl |
| Isku-dhafka Enzyme (Taq&M-MLV) | 1.5 μl |
| Aasaaska iyo Baarista | 1μL |
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).
