Multiplex qPCR Probe Premix
Cat Nama: HCB5051A
Ko te TaqMan multiplex qPCR Master Mix (Dye Based) he otinga-mua mo te 2 × te whakakaha PCR ine-wa-tui e tohuhia ana e te tairongo teitei me te motuhake, he puru te tae, me te whai hua o te taapiri tauira.Ko tenei hua he reagent 2× Whakaranu i mua i te whakaranu e taea ai te wha o nga tauhohenga PCR ine ine i roto i te puna tauhohenga kotahi.Kei roto i tenei hua te tikanga antibody kua whakarerekētia ki te wera-tīmatanga Taq whākōkī, tino whakapai ake i te tairongo whakakaha me te motuhake.I te wa ano, kua tino arotauhia e tenei hua te parepare tauhohenga-maha, ka taea te whakapai ake i te kaha o te whakakaha o te tauhohenga me te whakatairanga i te whakakaha whai hua o nga tauira iti-kutanga.Ka taea te whakamahi i tenei hua mo te taatai ine me te tātari ine maha.
Whakatakotoranga
| Tīmata wera | Hanga-i roto i te timatanga wera |
| Tikanga rapu | Ko te rapunga tuatahi |
| Tikanga PCR | qPCR |
| Polymerase | Taq DNA polymerase |
| Momo tauira | DNA |
Nga Tikanga Rokiroki
Ka tukuna te hua me te hukapapa maroke ka taea te rongoa ki te -25~ -15 ℃ mo te 2 tau.
Tohutohu
1. TauhohengaPūnaha
| Waehanga | Rōrahi (μL) | Te Whakamutunga Whakamutunga |
| 2× TaqMan multiplex qPCR Master Mix | 12.5 | 1× |
| Rahui tuatahi (10 μmol/L) a | × | 0.1 - 0.5 μmol/L |
| Ranu rangahau (10 μmol/L)b | × | 50 - 250 nmol/L |
| Tae tohutoro Rox | 0.5 | 1× |
| Te tauira DNA/cDNA | 1-10 | - |
| ddH2O | ki te 25 | - |
Tuhipoka:Whakaranuhia i mua i te whakamahi kia kore ai e nui nga mirumiru mai i te wiri kaha.
a.Te kukū tuatahi: He maha nga takirua o te Rakau Tuatahi, ko te nuinga o ia tuatahi i te kuhanga whakamutunga o te 0.2 μmol/L ka taea hoki te whakatika i waenga i te 0.1 me te 0.5 μmol/L ina tika.
b.Te kukū o te rangahau: Kei roto i te Probe Mix nga rangahau maha me nga tohu rereke rereke, a ka taea te whakatikatika i te kukū o ia tirotiro i waenga i te 50 me te 250 nmol/L i runga i nga ahuatanga motuhake.
1.Tohutoro tae Rox:Ka whakamahia i runga i nga taputapu whakakaha PCR Wā Tuturu penei i te Applied Biosystems ki te whakatika i te hapa o te tohu korikori i hangaia i waenga i nga puna;Ko tenei hua kaore he tohutoro tae Rox.Ka tūtohuhia te Cas#10200 ki te hiahiatia.
2.Te waimeha tauira: He tino tairongo te qPCR, a ka tūtohu kia waimeha i te tauira hei whakamahi.Mēnā he otinga kararehe cDNA te tauira, kaua e neke ake i te 1/10 o te rōrahi katoa.
3.Pūnaha tauhohenga: 25μL, 30μL, 50μL ranei e taunaki ana hei whakarite i te whai huatanga me te tukuruatanga o te whakakaha ira ira.
4.Whakaritenga Pūnaha: Whakapaia koa i roto i te tauera tino ma, ka whakamahi i nga tohutohu me nga ngongo hohenga kaore he toenga karihi;e taunaki ana kia whakamahia nga tohutohu me nga kaata tātari.A ape i te paheketanga whakawhiti me te poke aerosol.
2.Hotaka tauhohenga
| Huahi huringa | Temp. | Te wa | Nga huringa |
| Ko te whakataa-tuatahi | 95 ℃ | 5min | 1 |
| Te whakahekenga | 95 ℃ | 15 hēkona | 45 |
| Whakawhanaunga/Torohanga | 60 ℃ | 30 hēkona |
Tuhipoka:
1.Whakapiri/Torohanga: Ka taea te whakatika i te pāmahana me te wa i runga i te uara Tm tuatahi i hoahoatia.
2.Te whiwhi tohu tohu rererangi: He rereke te wa whiwhi tohu tohu rererangi e hiahiatia ana mo nga taputapu rapu qPCR rereke, me whakarite kia rite ki te wa iti.Ko te wa o etahi taputapu noa kua whakaritea e whai ake nei:
20 hēkona: Whakamahi Biosystems 7700, 7900HT, 7500 Tere
31 hēkona: Applied Biosystems 7300
32 hēkona: Applied Biosystems 7500
Notes
Tena koa mauhia nga PPE e tika ana, he koti taiwhanga me nga karapu, kia pai ai to hauora me to haumaru!
