Multiplex Ib Kauj Ruam RT-qPCR Premix-UNG
Cat No: HCB5145A
Multiplex One Step RT-qPCR Probe Kit (UDG Plus) yog cov khoom siv ntau yam PCR raws li RNA ua qauv.Nyob rau hauv tus txheej txheem ntawm kev sim, rov qab transcription thiab quantitative PCR tau ua nyob rau hauv tib lub raj, uas yooj yim rau kev sim ua hauj lwm thiab txo qhov kev pheej hmoo ntawm kev kis kab mob.Hauv cov khoom siv no, thawj strand cDNA tau ua haujlwm zoo los ntawm cov hluav taws kub-resistant Reverse Transcriptase thiab quantitatively amplified los ntawm HotStart Tag DNA Polymerase.Cov khoom siv feem ntau muaj cov tshuaj MP tsis zoo, cov enzymes sib xyaw, thiab lwm yam. Cov tshuaj tsis yog twb muaj Mg.2+thiab dNTP.Tsis tas li ntawd, cov yam ntxwv uas tuaj yeem cuam tshuam qhov tsis yog PCR amplification thiab txhim kho qhov kev ua kom zoo ntawm ntau qhov qPCR cov tshuaj tiv thaiv ntxiv, uas tuaj yeem ua kom muaj kev ua kom zoo dua qub thiab ua tiav ntau qhov kev tawm dag zog.Lub dUTP/UDG system tau ntxiv los tiv thaiv kev pheej hmoo ntawm cov kab mob aerosol.
Cheebtsam
1. Tsis pub
2. Enzyme Mix
Specification
| Pib kub | Built-in kub pib |
| Kev kuaj xyuas txoj kev | Primer-probe detection |
| PCR txoj kev | Ib kauj ruam RT-qPCR |
| Polymerase | Cov DNA polymerase |
| Hom qauv | DNA |
Kev cia khoom
Cov khoom xa tuaj nrog cov dej khov qhuav thiab tuaj yeem khaws cia ntawm -25 ~ -15 ℃ rau 1 xyoos.Nws yuav tsum tsis txhob nquagkhov-thaw.Nws raug pom zoo kom khaws cia nyias.
Cov lus qhia
1.Cov tshuaj tiv thaiv Qhov system
| Cheebtsam | Ntim (μL) | Thaum kawg Concentration |
| 2 × MP Buffer | 12.5 ib | 1 × |
| Enzyme Mix | 1 | - |
| Primer / Probe mix (2.5 μM) | 3 | 0,3m ua |
| Template RNA | 1-10 | - |
| RNase Dawb H2O | rau 25 | - |
Nco tseg:
Nco ntsoov sib tov zoo ua ntej siv, tsis txhob muaj npuas ntau dhau los ntawm kev vibration hnyav.
a.Primer concentration: Primer mix nrog rau multiplex primer, nyob ntawm qhov xwm txheej zoo primer concentration tej zaum ntawm 0.l thiab 1.0μM.
b.Kev sojntsuam concentration: Kev sojntsuam sib xyaw suav nrog multiplex sojntsuam sau npe sib txawv pawg fluorescent, nyob ntawm qhov xwm txheej zoo tshaj plaws kev sojntsuam concentration tej zaum ntawm 0.05 thiab 0.5μM.
c.Template dilution: qPCR yog qhov rhiab heev thiab nws raug pom zoo kom dilute lub template.Tus nqi tswj Ct yog tsim los ntawm 20 thiab 35.
d.Kev npaj: Thov npaj rau hauv lub rooj ua haujlwm huv huv, thiab cov raj xa dej thiab cov tshuaj tiv thaiv tsis muaj nuclease residue;nws raug pom zoo kom siv rab phom taub hau nrog cov khoom lim.Tsis txhob kis kab mob sib kis thiab aerosol paug.
2. Ua kom zoo caij tsheb kauj vabraws tu qauv
| Lub voj voog kauj ruam | Temp. | Sijhawm | Lub voj voog |
| Rov qab transcription | 50 ℃ib | 20 feeb | 1 |
| Thawj-denaturation | 95 ℃ | 5 feeb | 1 |
| Amplification cov tshuaj tiv thaiv | 95 ℃ | 15 sec |
40-45 : kuv |
| 60 ℃ b | 30 sec c |
Nco tseg:
a.Kev hloov pauv rov qab: Qhov kub tuaj yeem xaiv 42 ° C lossis 50 ° C.
b.Amplification cov tshuaj tiv thaiv: Qhov kub yog hloov raws li tus nqi Tm ntawm cov primers tsim.
c.Fluorescence teeb liab tau txais: Thov teeb qhov kev sim ua raws li qhov yuav tsum tau ua ntawm phau ntawv ntsuas ntsuas.
Sau ntawv
Thov hnav PPE uas tsim nyog, xws li lub tsho hauv chav kuaj thiab hnab looj tes, kom ntseeg tau tias koj txoj kev noj qab haus huv thiab kev nyab xeeb.
