RT-LAMP Midabaynta Masterka Isku-dhafka HCB5204A
Alaabtani waxay ka kooban tahay falcelinta falcelinta, RT-Enzymes Mix (Bst DNA polymerase and transcriptase reverse kulaylka), Ilaaliyeyaasha lyophilized iyo qaybaha dheeha koromojiinka.Si aad u isticmaasho, kaliya isticmaal Buffer, enzyme falcelinta iyo furaha waa la isku qasan yahay waxaana lagu daraa qaabka;ku darida ilaalinta lyophilized waxay noqon kartaa mid toosan.Waxay ku xidhnayd lyophilizer iyo lyophilized, oo kaliya furayaasha iyo qaababka ayaa lagu daray markii la isticmaalay.Xirmadani waxa ay bixisa ogaysiin muuqaal ah oo degdeg ah oo cad oo xoojin ah, taas oo falcelinta taban lagu muujiyay casaanka iyo falcelinta togan ee lagu muujiyay isbeddelka jaalaha ah.
Qayb
| Qayb | HCB5204A-01 | HCB5204A-02 | HCB5204A-03 |
| Kordhinta Loop-dhexdhexaadiyay (oo leh dheeh) | 0.96 ml | 4.80ml×2 | 9.60 ml×10 |
| RT-Enzymes Isku-dhafka | 270 μL | 2.70 ml | 2.70 ml×10 |
| Difaaca lyophilized | 0.96 ml×2 | 9.60ml×2 | 9.60ml×20 |
Codsiyada
Si loo kordhiyo DNA-ga ama RNA isothermal.
Xaaladaha Kaydinta
Lagu raray baraf qallalan, oo lagu kaydiyay -25 ~ -15 ℃.Iska ilaali baraf-dhalaalid joogto ah, alaabtu waxay shaqaynaysaa 12 bilood.
Baratakoolka
1.Ku dhalaali furaha falcelinta si loogu isticmaalo heerkulka qolka.Vortex si kooban ama u rogo tuubooyinka dhawr jeer si aad si fiican isugu walaaqdo, ka dibna sentrifuge si aad dareeraha ugu ururiso xagga hoose ee tuubada.
2.Diyaarinta habka falcelinta.Reagent-kan waxaa lagu diyaarin karaa laba hab oo falcelineed, isku darka falcelinta dareeraha ah iyo isku dhafka nidaamka lyophilized.
1) Diyaari isku dhafka falcelinta dareeraha
| Qayb | Mugga |
| Kordhinta Loop-dhexdhexaadiyay (oo leh dheeh) | 10 μl |
| RT-Enzymes Isku-dhafka | 2.8 μl |
| 10 × Isku dhafka koowaada | 5 μL |
| Qaababka DNA/RNA b | × μL |
| Biyo aan Nukliyeer lahayn | Ilaa 50 μl |
2) Isku darka nidaamka Lyophilization
① Diyaari isku darka lyophilized
| Qayb | Mugga |
| Kordhinta Loop-dhexdhexaadiyay (oo leh dheeh) | 10 μl |
| Difaaca lyophilized | 20 μL |
| RT-Enzymes Isku-dhafka | 2.8 μl |
| Biyo aan Nukliyeer lahayn | Ilaa 50 μl |
② Lyophilization: Isku-darka la diyaariyey waxaa lagu lyophilized nidaamka 50μL
③ Diyaarso isku darka falcelinta
| Qayb | Mugga |
| Isku darka Lyophilized | 1 gabal |
| 10 × Isku dhafka koowaada | 5 μL |
| Qaababka DNA/RNA b | × μL |
| Biyo aan Nukliyeer lahayn | Ilaa 50 μl |
Xusuusin:
1) ah.10×Primer Mix : 16 μM FIP/BIP, 2 μM F3/B3, 4 μM Loop F/B;
2) b.DEPC (biyaha ku milma) ayaa lagula talinayaa templ nucleic acid.
1.Ku dhaji 65°C ilaa 30-45mins, kaas oo si habboon loo kordhin karo iyadoo loo eegayo isbeddelka midabka wakhtiga falcelinta.
2.Sida laga soo xigtay isha qaawan, jaalaha ayaa ahaa mid togan, casaankuna wuxuu ahaa mid xun.
Xusuusin
1.milixdu waxa ay ka soo muuqan kartaa gunta hoose ee tuubada, vortex si kooban ama tuubooyinka roganaya dhawr jeer si si fiican loogu qaso heerkulka qolka.
2.Heerkulka falcelinta waxaa lagu hagaajin karaa inta u dhaxaysa 62 ℃ iyo 68 ℃ iyadoo loo eegayo xaaladda asaasiga ah.
3.Dib-u-hawlgalayaasha baakadaysan waa in aanay hawada gelin muddo dheer.
4.Dareen-celinta casaanka iyo jaalaha ah waxay ku xiran tahay isbeddelka pH ee nidaamka falcelinta, fadlan ha isticmaalin ka kooban xalka kaydinta Tris nucleic acid, oo lagu taliyay in la isticmaalo ddH2Aashitada nucleic ee kaydsan;
5.Tijaabada waa in la jaangooyaa, oo ay ku jiraan diyaarinta habka falcelinta, lyophilization, iyo habraaca muunad samaynta iyo muunad kordhinta;
6.Si looga fogaado wasakheynta, waxaa lagu talinayaa in lagu diyaariyo habka falcelinta ee kursiga keydka aadka u nadiifka ah, kuwa kale Ku dar qaabyaalo daboolka qiiqa ee qolka si looga fogaado faragelin been ah.
