Multiplex One Step RT-qPCR Premix-UNG
Cat Nama: HCB5145A
Multiplex One Step RT-qPCR Probe Kit (UDG Plus) he kete PCR ine ine maha i runga i te RNA hei tauira.I roto i te tukanga o te whakamatautau, i mahia te whakamaoritanga whakamuri me te PCR ine i roto i te ngongo kotahi, i ngawari ai te mahi whakamatautau me te whakaiti i te tupono o te poke.I roto i tenei kete, i whakahiatotia te whenu tuatahi cDNA e te Reverse Transcriptase aatete wera me te whakanui ine e HotStart Tag DNA Polymerase.Kei roto i te kete te nuinga o te kaipara MP kua pai ake, te ranunga o nga enzymes, me era atu2+me te dNTP.I tua atu, ko nga mea e kaha ana ki te whakakore i te kaha o te PCR kore-motuhake me te whakapai ake i te kaha o te kaha o te maha o nga tauhohenga qPCR ka taapirihia, ka taea te whakarite i te kaha o te whakakaha me te kawe ki te tauhohenga whakakaha maha.I taapirihia te punaha dUTP/UDG kia pai ai te aukati i te kino o te aerosol.
Waehanga
1. Kaipupuri
2. Whakaranu Enzyme
Whakatakotoranga
| Tīmata wera | Hanga-i roto i te timatanga wera |
| Tikanga rapu | Ko te rapunga tuatahi |
| Tikanga PCR | Kotahi te taahiraa RT-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 1 tau.Me karo tonuwhakatio-rewa.E taunaki ana kia penapena motuhake.
Tohutohu
1.Tauhohenga Pūnaha
| Waehanga | Rōrahi (μL) | Te Whakamutunga Whakamutunga |
| 2 × MP Kaipupuri | 12.5 | 1× |
| Whakaranu Enzyme | 1 | - |
| Raupapa tuatahi/Matai (2.5 μM) | 3 | 0.3μM |
| RNA tauira | 1-10 | - |
| RNase Free H2O | ki te 25 | - |
Tuhipoka:
Kia pai te whakauru i mua i te whakamahi, karohia nga mirumiru nui na te wiri tutu.
a.Te kukū tuatahi: Te ranunga tuatahi tae atu ki te maha o te tuatahi, i runga i te ahuatanga tino pai te kukū tuatahi i waenga i te 0.l me te 1.0μM.
b.Te kukū o te rangahau: Te ranunga rangahau tae atu ki te roopu rekoata tapanga tapanga maha, i runga i te ahua o te kukū rangahau tino pai pea kei waenga i te 0.05 me te 0.5μM.
c.Te waimeha tauira: He tino tairongo te qPCR a ka tūtohu kia waimeha i te tauira.He pai te uara Ct mana i waenga i te 20 me te 35.
d.Whakaritenga Pūnaha: Whakapaia koa i roto i te tepu mahi ultra ma, me te pipettor me te ngongo hohenga kaore he toenga karihi;e taunaki ana kia whakamahia te upoko pu me te huānga tātari.A ape i te paheketanga whakawhiti me te poke aerosol.
2. Paihikara kua arotauKawa
| Porohita takahanga | Temp. | Te wa | Nga huringa |
| Whakamuri whakamuri | 50 ℃a | 20 meneti | 1 |
| Ko te whakataa-tuatahi | 95℃ | 5min | 1 |
| Tauhohenga whakakaha | 95℃ | 15 hēkona |
40-45 |
| 60 ℃ b | 30 hēkona c |
Tuhipoka:
a.Whakamuri whakamuri: Ka taea e te pāmahana te 42°C, 50°C ranei.
b.Tauhohenga Whakanuia: Ka whakatikahia te pāmahana kia rite ki te uara Tm o nga papa tuatahi i hangaia.
c.Te whiwhi tohu ororongo: Whakatakotoria te tikanga whakamatautau kia rite ki nga whakaritenga o te pukapuka taputapu.
Notes
Tena koa mau nga PPE e tika ana, penei i te koti taiwhanga me nga karapu, kia pai ai to hauora me to haumaru.
