Multiplex qPCR Probe Premix
Cat No: HCB5051A
TaqMan multiplex qPCR Master Mix (Dye Based) yog ib qho kev daws teeb meem ua ntej rau 2 × real-time quantitative PCR amplification tus yam ntxwv los ntawm siab rhiab heev thiab tshwj xeeb, uas yog xiav xim, thiab muaj cov nyhuv ntawm cov qauv ntxiv.Cov khoom no yog 2 × Mix pre-mixed reagent uas ua rau kom txog li plaub lub fluorescent quantitative PCR cov tshuaj tiv thaiv hauv ib qho tshuaj tiv thaiv zoo.Cov khoom no muaj cov tshuaj tiv thaiv kab mob hloov pauv hloov mus rau qhov kub-pib Taq enzyme, zoo heev txhim kho lub amplification rhiab heev thiab tshwj xeeb.Nyob rau tib lub sijhawm, cov khoom no tau ua kom zoo tshaj plaws ntawm ntau qhov tsis sib haum xeeb, uas tuaj yeem txhim kho qhov kev ua kom zoo ntawm cov tshuaj tiv thaiv thiab txhawb kom muaj txiaj ntsig zoo ntawm cov qauv qis qis.Cov khoom no tuaj yeem siv rau genotyping thiab multiplex quantitative tsom xam.
Specification
| Pib kub | Built-in kub pib |
| Kev kuaj xyuas txoj kev | Primer-probe detection |
| PCR txoj kev | 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 2 xyoos.
Cov lus qhia
1. Kev tshwm simQhov system
| Cheebtsam | Ntim (μL) | Thaum kawg Concentration |
| 2 × TaqMan multiplex qPCR Master Mix | 12.5 ib | 1 × |
| Primer sib tov (10 μmol / L) a | × | 0.1-0.5 μmol / L |
| Kev sojntsuam sib xyaw (10 μmol / L)b | × | 50-250 nmol / L |
| Rox siv dye | 0.5 | 1 × |
| Template DNA/cDNA | 1-10 | - |
| ddH ua2O | mus 25 | - |
Nco tseg:Sib tov kom huv si ua ntej siv kom tsis txhob muaj npuas ntau dhau los ntawm kev tuav lub zog.
a.Primer concentration: Primer Mix muaj ntau khub ntawm primers, feem ntau txhua tus primer ntawm qhov kawg concentration ntawm 0.2 μmol / L thiab tuaj yeem hloov kho ntawm 0.1 thiab 0.5 μmol / L raws li qhov tsim nyog.
b.Kev sojntsuam concentration: Kev sojntsuam sib xyaw muaj ntau qhov kev sojntsuam nrog cov cim fluorescence sib txawv, thiab qhov concentration ntawm txhua qhov kev sojntsuam tuaj yeem hloov kho ntawm 50 thiab 250 nmol / L raws li qhov xwm txheej tshwj xeeb.
1.Rox dye reference: Nws yog siv ntawm Real Time PCR amplification instrument xws li Applied Biosystems los kho qhov yuam kev ntawm fluorescence teeb liab generated ntawm qhov dej;Cov khoom no tsis muaj Rox dye reference.Cas #10200 raug pom zoo yog tias xav tau.
2.Template dilution: qPCR yog qhov rhiab heev, thiab nws raug nquahu kom dilute tus qauv siv.Yog tias tus qauv yog cDNA cov khoom lag luam, cov qauv ntim yuav tsum tsis pub tshaj 1/10 ntawm tag nrho cov ntim.
3.Cov tshuaj tiv thaiv kab mob: 25μL, 30μL lossis 50 μL raug pom zoo los xyuas kom meej qhov ua tau zoo thiab rov ua dua ntawm lub hom phiaj gene amplification.
4.Kev npaj cov txheej txheem: Thov npaj rau hauv lub rooj zaum huv si, thiab siv cov lus qhia thiab cov tshuaj tiv thaiv kab mob tsis muaj nuclease residue;nws raug pom zoo kom siv cov lus qhia nrog lim cartridges.Tsis txhob kis kab mob sib kis thiab aerosol paug.
2.Reaction program
| Lub voj voog kauj ruam | Temp. | Sijhawm | Lub voj voog |
| Thawj-denaturation | 95 ℃ | 5 feeb | 1 |
| Denaturation | 95 ℃ | 15 sec | 45 |
| Annealing / Extension | 60 ℃ | 30 sec |
Nco tseg:
1.Annealing / Extension: Qhov kub thiab lub sijhawm tuaj yeem hloov kho kom haum raws li tus nqi tsim primer Tm.
2.Fluorescence signal acquisition: Lub sij hawm fluorescence tau txais cov teeb liab yuav tsum tau rau cov cuab yeej nrhiav kom pom qPCR sib txawv, thov teem caij raws li lub sijhawm tsawg kawg nkaus.Lub sijhawm ntawm ntau yam twj paj nruag yog teem raws li hauv qab no:
20 sec: Siv Biosystems 7700, 7900HT, 7500 Ceev
31 sec: Siv Biosystems 7300
32 sec: Siv Biosystems 7500
Sau ntawv
Thov hnav PPE tsim nyog, xws li lub tsho tiv no thiab hnab looj tes, txhawm rau ua kom koj noj qab haus huv thiab nyab xeeb!
