RT-LAMP Colormetric Master Ivanga HCB5204A
Iki gicuruzwa kirimo reaction ya reaction, RT-Enzymes ivanze (Bst ADN polymerase na Bris DNA irwanya ubushyuhe), porotokoro ya lyofilize hamwe nibigize irangi rya chromogene.Gukoresha, koresha gusa Buffer, reaction ya enzyme na primer bivanze kandi byongewe kubishusho;wongeyeho lyofilized protekant irashobora kuba igororotse.Yahujwe na lyophilizer na lyofilize, kandi primers na templates gusa byongeweho mugihe byakoreshejwe.Iki gikoresho gitanga byihuse, bisobanutse neza byerekana amplification, iyo reaction mbi igaragarira mumutuku kandi reaction nziza igaragazwa no guhindura umuhondo.
Ibigize
| Ibigize | HCB5204A-01 | HCB5204A-02 | HCB5204A-03 |
| Umuyoboro uhuza Amplification Buffer (ufite irangi) | 0,96 mL | 4.80 mL × 2 | 9.60 mL × 10 |
| RT-Enzymes ivanze | 270 μL | 2.70 mL | 2.70 mL × 10 |
| Kurinda Lyophilized | 0,96 mL × 2 | 9.60 mL × 2 | 9.60 mL × 20 |
Porogaramu
Kuri ADN cyangwa RNA isothermal amplification.
Ububiko
Gutwarwa na ice cyumye, kibitswe kuri -25 ~ -15 ℃.Irinde gukonjesha kenshi, ibicuruzwa bifite agaciro mumezi 12.
Porotokole
1.Kuramo buffer reaction kugirango ikoreshwe mubushyuhe bwicyumba.Vortex muri make cyangwa ihindure imiyoboro inshuro nyinshi kugirango ivange neza, hanyuma centrifuge kugirango ikusanyirize amazi munsi yigituba.
2.Gutegura sisitemu yo kwitwara.Iyi reagent irashobora gutegurwa muburyo bubiri bwa reaction, kuvanga ibintu bivanze no kuvanga sisitemu.
1) Tegura kuvanga ibintu bivanze
| Ibigize | Umubumbe |
| Umuyoboro uhuza Amplification Buffer (ufite irangi) | 10 μL |
| RT-Enzymes ivanze | 2.8 μL |
| 10 mix Primer mixa | 5 μL |
| Inyandikorugero ADN / RNA b | × μL |
| Amazi adafite ingufu | Kugera kuri 50 μL |
2) Sisitemu yo kuvanga Lyophilisation
Tegura kuvanga lyofilize
| Ibigize | Umubumbe |
| Umuyoboro uhuza Amplification Buffer (ufite irangi) | 10 μL |
| Kurinda Lyophilized | 20 μL |
| RT-Enzymes ivanze | 2.8 μL |
| Amazi adafite ingufu | Kugera kuri 50 μL |
Oph Lyophilisation: Ivanga ryateguwe ryakozwe muri sisitemu ya 50μL
Tegura kuvanga reaction
| Ibigize | Umubumbe |
| Kuvangavanga | Igice 1 |
| 10 mix Primer mixa | 5 μL |
| Inyandikorugero ADN / RNA b | × μL |
| Amazi adafite ingufu | Kugera kuri 50 μL |
Inyandiko:
1) a.10 × Primer mix: 16 μM FIP / BIP, 2 μM F3 / B3, 4 μM Umuzingi F / B;
2) b.DEPC (soluble water) irasabwa kuri nucleic aside templ.
1.Incubate kuri 65 ° C kuri 30-45mins, irashobora kongerwa uko bikwiye ukurikije ihinduka ryibara ryigihe.
2.Ukurikije ijisho ryonyine, umuhondo wari mwiza naho umutuku ukaba mubi.
Inyandiko
1.Umunyu urashobora kugaragara munsi yigituba cya buffer, vortex mugihe gito cyangwa guhinduranya imiyoboro inshuro nyinshi kugirango uvange neza mubushyuhe bwicyumba.
2.Ubushyuhe bwa reaction bushobora kuba bwiza hagati ya 62 ℃ na 68 ℃ ukurikije imiterere ya primers.
3.Ibikoresho byapakiwe ntibigomba guhura numwuka igihe kirekire.
4.Ibara ry'umutuku n'umuhondo biterwa na pH ihinduka rya sisitemu ya reaction, nyamuneka ntukoreshe igisubizo kibitse cya Tris nucleic aside, gisabwa gukoresha ddH2O acide nucleic yabitswe;
5.Ubushakashatsi bugomba kuba busanzwe, harimo gutegura sisitemu yo kubyitwaramo, lyophilisation, hamwe no gutunganya icyitegererezo hamwe no kongera icyitegererezo;
6.Kugira ngo wirinde kwanduza, birasabwa gutegura sisitemu yo kubyitwaramo mu ntebe isukuye cyane, mu zindi Ongeraho inyandikorugero kuri fume hood yicyumba kugirango wirinde kwivanga nabi.
