Hal Tilaabo RT-qPCR SYBR Cagaar Premix
Bisadda No: HCB5140A
Hal Tilaabo RT-qPCR Syber Cagaar Premix waxaa loogu talagalay qiyaasta fluorescence ee ku salaysan SYBR Cagaaran I dheeha.Isticmaalka aasaasayaasha hidde-sidaha gaarka ah, qoraal-ku-celinta iyo falcelinta qPCR ayaa lagu dhammeeyaa hal tubo, taas oo meesha ka saaraysa baahida loo qabo fur-furidda iyo dhuumaha soo noqnoqda, taasoo si weyn u wanaajisa hufnaanta qiimaynta iyo yaraynta khatarta wasakhowga.Shaybaarada RNA, qalabku waxa uu u shaqeeyaa Reverse Transcriptase kulayl u adkaysta si ay u abuurto cDNA hufan iyo HotStart Taq DNA Polymerase si ay u kordhiso tiradaMarka la eego nidaamka kaydinta la hagaajiyay, dareenka xirmadu wuxuu noqon karaa mid aad u sarreeya ilaa 0.1 pg ee bartilmaameedyada aadka loo muujiyay iyo ilaa 1 pg ee bartilmaameedyada si dhexdhexaad ah loo muujiyay.Qalabku wuxuu ku habboon yahay kordhinta iyo qiyaasidda shaybaarka DNA-ga.Waxay awood u siinaysaa ogaanshaha xasaasiga ah iyo qiyaasidda asiidhyada nucleic-ka ah ee laga soo qaaday muunado dhir iyo xayawaan oo kala duwan, unugyo iyo noolayaal yaryar.
Qaybaha
| No | Magaca | Mugga | Mugga |
| 1 | Buffer Sare | 250 μL | 2×1.25 mL |
| 2 | Isku-dhafka Enzyme Sare | 20 μL | 200 μL |
| 3 | RNase bilaash ah H2O | 250 μL | 2×1.25 mL |
Xaaladaha Kaydinta
Alaabtan waa in lagu kaydiyaa meel -25~-15℃ ka fog iftiinka muddo 1 sano ah.
Tilmaamaha
1.Reaction habaynta nidaamkad
| Qaybaha | Mugga (μL) | Mugga (μL) | U fiirsashada kama dambaysta ah |
| Buffer Sare | 12.5 | 25 | 1× |
| Isku-dhafka Enzyme Sare | 1 | 2 | - |
| Horudhac hore (10 μmol/L)a | 0.5 | 1 | 0.2 μmol/L |
| Dib-u-celinta hore (10 μmol/L)a | 0.5 | 1 | 0.2 μmol/L |
| RNA Tamplateb | X | X | - |
| RNase bilaash ah H2Oc | ilaa 25 | ilaa 50 | - |
Xusuusin:
1) ah.Tfiirsiga ugu dambeeya ee asaasiga ahi wuxuu ahaa 0.2 μmol/L, kaas oo sidoo kale lagu hagaajin karo inta u dhaxaysa 0.1 iyo 1μmol/L sida ku habboon.
2) b.Reagent waa mid aad xasaasi u ah, oo leh wadarta guud ee RNA inta u dhaxaysa 1pg-1μg, iyo tijaabinta shaybaarada bini'aadamka waxay muujisay gelinta ugu fiican ee 1 pg-100 ng, xakamaynta qiimaha guud ee Ct ee inta u dhaxaysa 15-30 sida ku habboon.
3) c.Waxaa lagu talinayaa in la isticmaalo 20μL ama 50μL si loo hubiyo ansaxnimada iyo soo saaridda kordhinta hiddo-wadaha bartilmaameedka ah.
4) d.Fadlan ku diyaari kursiga keydka aadka u nadiifka ah oo isticmaal talooyinka iyo tuubooyinka falcelinta ee ka xorta ah nuclease;talooyinka leh kartoonada shaandhada ayaa lagula talinayaa.Ka fogow wasakhowga isdhaafka ah iyo wasakhowga hawada.
2.Barnaamijka falcelinta
| Tallaabo wareeg | Heerkulka | Waqtiga | Wareegtada |
| Dib u qorista | 50℃a | 6 daqiiqo | 1 |
| Denaturation bilowga ah | 95℃ | 5 daqiiqo | 1 |
| Falcelinta weynaynta | 95℃ | 15 ilbiriqsi | 40 |
| 60℃b | 30 ilbiriqsi | ||
| Marxaladda qalooca dhalaalaysa | Qalab Default | 1 | |
Xusuusin:
1) ah.Heerkulka beddelka qoraalka ayaa la dooran karaa inta u dhaxaysa 50-55 ° C iyadoo loo eegayo baahida tijaabada ah.Shaybaarada DNA-da, habka dib-u-qorista waa laga tagi karaa.
2) b.Xaalado gaar ah heerkulka nuugista/kordhinta waxaa lagu hagaajin karaa iyadoo loo eegayo qiimaha asaasiga ah ee Tm, 60°C ayaa lagula talinayaa.
Xusuusin
1. Alaabtan waxaa loogu talagalay isticmaalka cilmi-baarista oo keliya.
2. Fadlan ku shaqee jaakadaha shaybaadhka iyo galoofyada la tuuri karo, badbaadadaada awgeed.
