RT-LAMP Colormetric (Umupira wa Lyophilized)
Iki gicuruzwa kirimo reaction ya reaction, RT-Enzymes ivanze (Bst ADN polymerase ya Bst na anti-transcriptase irwanya ubushyuhe), kurinda lyofilize hamwe nibigize irangi rya chromogenic.Igicuruzwa ni ubwoko bwumupira wamaguru, ukoresheje gusa primers na templates.Iki gikoresho gitanga byihuse, bisobanutse neza byerekana amplification, iyo reaction mbi igaragarira mumutuku kandi reaction nziza igaragazwa no guhindura umuhondo.
Ibigize
RT-LAMP Colormetric Master mix (Isaro rya Lyophilized)
Porogaramu
Kuri ADN cyangwa RNA isothermal amplification.
Ububiko
Gutwarwa no kubikwa kuri 2 ~ 8 ℃.Igicuruzwa gifite agaciro mumezi 12.
Porotokole
1.Kuramo umubare uhuye na powder ya Lyophilized ukurikije umubare wibizamini.
2.Tegura kuvanga reaction
| Ibigize | Umubumbe |
| RT-LAMP Colormetric Master mix (Isaro rya Lyophilized) | Igice 1 (amasaro 2) |
| 10 mix Primer mixa | 5 μL |
| Inyandikorugero ADN / RNA b | 45 μL |
Inyandiko:
1. 10 × Kwishyira hamwe kwa Primer: 16 μM FIP / BIP, 2 μM F3 / B3, 4 μM Umuzingi F / B;
2. Nucleic aside templates irasabwa gushonga ukoresheje amazi ya DEPC.
3.Incubate kuri 65 ° C kuri 30-45mins, irashobora kongerwa uko bikwiye ukurikije ihinduka ryibara ryigihe.
4.Ukurikije ijisho ryonyine, umuhondo wari mwiza naho umutuku ukaba mubi.
Inyandiko
1.Ubushyuhe bwa reaction bushobora kuba bwiza hagati ya 62 ℃ na 68 ℃ ukurikije imiterere ya primer.
2.Ibikoresho byapakiwe ntibigomba guhura numwuka igihe kirekire.
3.Ibara ritukura n'umuhondo biterwa na pH ihinduka rya sisitemu ya reaction, nyamuneka ntukoreshe igisubizo kirimo ububiko bwa Tris nucleic aside, busabwa gukoresha ddH2O aside nucleic yabitswe.
4.Ubushakashatsi bugomba gukorwa muburyo busanzwe, harimo gutegura sisitemu yo kubyitwaramo, kuvura icyitegererezo no kongeramo icyitegererezo.
5.Birasabwa gutegura sisitemu yo kwitwara mumeza ya ultra-isuku hanyuma ukongeramo inyandikorugero muri fume hood yibindi byumba kugirango wirinde ibinyoma.
