Multiplex One Step RT-qPCR Premix
Whakaahuatanga
Cat Nama: HCR5141A
Ko te Multiplex One Step RT-qPCR Premix he kete PCR ine ine i runga i te RNA hei tauira.I roto i te whakamatautau, ka mahia te whakamaoritanga whakamuri me te PCR ine i roto i te ngongo tauhohenga kotahi, ka ngawari te mahi whakamatautau me te whakaiti i te tupono o te poke.Ka taea te whakamahi i te hoahoa ahurei o te whakahiato me te whakauru i roto i te punaha lyophilized kotahi-taahiraa.Ka whakamahia e te kete te whakamuri Transcriptase whakamuri wera mo te whakahiatotanga o te whenu tuatahi cDNA ma te whakamahi i te Hotstart Taq DNA Polymerase mo te whakanui ine.Kei roto he parepare tauhohenga tino pai, he ranunga o nga enzymes me etahi atu mea, me nga mea e aukati ana i te whakakaha PCR kore-motuhake me te whakarei ake i te kaha whakakaha o nga tauhohenga qPCR maha i taapirihia, e taea ai te whakakaha ine ine rewharewha maha i te wa e whakarite ana i te kaha o te whakakaha o nga tuatahi.
Waehanga
| Ingoa |
| 1. Lyo-Buffer |
| 2. Lyo-Enzyme Mix |
| 3. Lyo kaitiaki |
Tikanga wakakatote
A: Lyo-buffer me te kaitiaki: -25 ~ -15 ℃, ko te 1 tau te ora o te papa.
B: Te ranunga Lyo-enzyme, 2-8 ℃, e 6 marama te roa o te ora.
Tohutohu mo te mahi
1. Pūnaha Tauhohenga (Tangohia te 25μL hei tauira)
| Waehanga | Rōrahi (μL) | Te Whakamutunga Whakamutunga |
| Lyo-Buffer | 6 | 1* |
| Lyo-Enzyme Mix | 1 | - |
| Lyo-Kaitiaki | 8 | - |
| Whakaranu Tuatahi (10μM) | 1 | 0.1- 1uM |
| Whakaranu Matau (10μM) | 0.5 | 0.05-0.5uM |
| Tātauira RNA | 5 | - |
| DEPC H2O | Tae atu ki te 25 | - |
2. Kawa Paihikara Arotau
1) Kawa Paihikara Paerewa
| Te wahanga tauhohenga | Te pāmahana | Te wa | Porohita | |
| 1 | Whakamuri whakamuri | 50°Ca | 10 min | 1 |
| 2 | Te whakahekenga tuatahi | 95°C | 5min | 1 |
| 3 | Tauhohenga whakakaha | 95°C | 15 hēkona | 45 huringa |
| 60°Cb | 30 hēkonac |
2) Kawa Paihikara Tere
|
| Te wahanga tauhohenga | Te pāmahana | Te wa | Porohita |
| 1 | Whakamuri whakamuri | 50°Ca | 2min | 1 |
| 2 | Te whakahekenga tuatahi | 95°C | 2 hēkona | 1 |
| 3 | Tauhohenga whakakaha | 95°C | 1 hēkona |
45 huringa |
| 60°Cb | 13 hēkonac |
Tuhipoka:
a) Whakamuri whakamaoritanga: Ka taea e te pāmahana te kowhiri i te 42°C ranei 50°C mo te 10-15 meneti.
b) Tauhohenga whakakaha: Ka whakatikahia te pāmahana i runga i te uara Tm o nga papa tuatahi i hangaia.
c)Korewharewha tohu rironga: Whakatakotoria te tikanga whakamatautau kia rite ki nga whakaritenga o tepukapuka taputapu.
Nga Korero Hangarau/Whakaahuatanga
| Tīmata wera | Hanga-i roto i te timatanga wera |
| Tikanga rapu | Ko te rapunga tuatahi |
| Tikanga PCR | Kotahi te taahiraa RT-qPCR |
| Momo tauira | RNA |
Notes
1. Ko tenei hua mo te rangahau anake.
2. Tena koa mauhia nga PPE e tika ana, he koti taiwhanga me nga karapu, hei whakarite i to hauora me to haumaru！
