Kotahi Hipanga Tere RT-qPCR Probe Premix-UNG
Cat Nama: HCR5143A
One Step RT-qPCR Probe Kit (MĀ TE KAUPAPA) he kete rapunga tere RT-qPCR e tika ana mo te PCR ine ine kotahi-plex ranei ma te whakamahi i te RNA hei tauira (penei i te huaketo RNA).Ka whakamahia e tenei hua te reanga hou o te Taq DNA Polymerase kua whakarerekehia e te antibody me te Reverse Transcriptase kotahi-taahiraa i whakatapua, me te papaa pai mo te whakakaha tere, he tere ake te tere o te whakakaha, he pai ake te whakakaha me te motuhake.Ka tautokohia te whakakaha taurite i roto i te kotahi-plex me te multiplex o nga tauira kukū iti me te teitei i roto i te wa poto.
Waehanga
1. 5×RT-qPCR pūreirei (U+)
2. Rahui Enzyme (U+)
Tuhipoka:
a.5×RT-qPCR buffer (U+)kei roto te dNTP me te Mg2+;
b.Ko te ranunga Enzyme (U+) kei roto ko te whakamuri transcriptase, Hot Start Taq DNA polymerase, te aukati RNase me te UDG;
c.Whakamahia nga tohutohu RNase-Free, ngongo EP, etc.
I mua i te whakamahi, tino whakaranuhia te 5×RT-qPCR buffer (U+).Mēnā he rerenga i muri i te whakamaroketanga, tatari kia hoki mai te parepare ki te mahana o te rūma, uruhia ka memeha, katahi ka whakamahia noa.
Nga Tikanga Rokiroki
Ka tukuna te hua me te hukapapa maroke ka taea te rongoa ki te -25~ -15 ℃ mo te 1 tau.
Tohutohu
1. Tauhohenga Pūnaha
| Waehanga | Rōrahi (20 μL tauhohenga) |
| 2 ×RT-qPCR parepare | 4μL |
| Rahui Enzyme (U+) | 0.8μL |
| Primer Whakamua | 0.1~ 1.0μM |
| Tuatahi Whakamuri | 0.1~ 1.0μM |
| TaqMan Probe | 0.05~0.25μM |
| tauira | X μL |
| RNase-Kore Wai | ki te 25μL |
Tuhipoka: Ko te 10-50μL te rōrahi tauhohenga.
2. Kawa Paihikara (Stohu)
| Porohita takahanga | Temp. | Te wa | Nga huringa |
| Whakamuri Whakamuri | 55 ℃ | 10 min | 1 |
| Whakahekenga Tuatahi | 95 ℃ | 30 hēkona | 1 |
| Te whakahekenga | 95 ℃ | 10 hēkona | 45 |
| Te whakawhanaunga/Torohanga | 60 ℃ | 30 hēkona |
| Kawa Paihikara (Tere) Porohita takahanga |
Temp. |
Te wa |
Nga huringa |
| Whakamuri Whakamuri | 55 ℃ | 5 min | 1 |
| Whakahekenga Tuatahi | 95 ℃ | 5 s | 1 |
| Te whakahekenga | 95 ℃ | 3 s | 43 |
| Te whakawhanaunga/Torohanga | 60 ℃ | 10 s |
Tuhipoka:
a.Ko te pāmahana tuhi whakamuri kei waenga i te 50 ℃ ki te 60 ℃, ko te whakanui ake i te pāmahana ka awhina ki te whakanui i nga hanganga matatini me nga tauira ihirangi CG teitei;
b.Ko te pāmahana whakaenee tino pai me whakatika i runga i te uara Tm o te tuatahi, me te kowhiri i te waa poto rawa mo te kohinga tohu rewharewha i runga i te taputapu PCR Wā Tuturu.
Notes
Tena koa mauhia nga PPE e tika ana, he koti taiwhanga me nga karapu, kia pai ai to hauora me to haumaru!
