Ib Kauj Ruam RT-qPCR SYBR Green Premix
Cat No: HCB5140A
Ib Kauj Ruam RT-qPCR Syber Green Premix yog rau fluorescence kom muaj nuj nqis raws li SYBR Green I dye.Siv cov noob tshwj xeeb primers, thim rov qab thiab qPCR cov tshuaj tiv thaiv tau ua tiav nyob rau hauv ib lub raj, tshem tawm qhov xav tau rov qhib lub hau thiab cov raj xa dej, txhim kho kev soj ntsuam zoo thiab txo qhov kev pheej hmoo ntawm kev sib kis.Rau cov qauv RNA, cov khoom siv ua haujlwm kub-resistant Reverse Transcriptase rau cDNA synthesis thiab HotStart Taq DNA Polymerase rau kom muaj nuj nqis amplification.Nyob rau hauv lub optimized buffer system, qhov rhiab heev ntawm cov khoom siv yuav siab li 0.1 pg rau cov hom phiaj qhia tau zoo thiab siab li 1 pg rau cov hom phiaj me me.Cov khoom siv tsim nyog rau kev nthuav dav thiab kom muaj nuj nqis ntawm DNA kuaj.Nws tso cai rau kev tshawb nrhiav rhiab thiab kom muaj nuj nqis ntawm nucleic acids los ntawm cov qauv cog thiab tsiaj sib txawv, hlwb thiab kab mob.
Cheebtsam
| No | Lub npe | Ntim | Ntim |
| 1 | Advanced Buffer | 250 mL | 2 × 1.25 mL |
| 2 | Advanced Enzyme Mix | 20 mL | 200 mL |
| 3 | RNase Dawb H2O | 250 mL | 2 × 1.25 mL |
Kev cia khoom
Cov khoom no yuav tsum khaws cia ntawm -25 ~ -15 ℃ deb ntawm lub teeb rau 1 xyoos.
Cov lus qhia
1.Reaction system configurationd
| Cheebtsam | Ntim (μL) | Ntim (μL) | Thaum kawg concentration |
| Advanced Buffer | 12.5 ib | 25 | 1 × |
| Advanced Enzyme Mix | 1 | 2 | - |
| Forward Primer (10 μmol / L)a | 0.5 | 1 | 0.2 μmol / L |
| Rov qab Primer (10 μmol / L)a | 0.5 | 1 | 0.2 μmol / L |
| RNA Tamplateb | X | X | - |
| RNase Dawb H2Oc | rau 25 | rau 50 | - |
Nco tseg:
1) a.TNws qhov kawg primer concentration yog 0.2 μmol / L, uas tuaj yeem hloov kho ntawm 0.1 thiab 1μmol / L raws li qhov tsim nyog.
2) ib.Cov reagent yog qhov rhiab heev, nrog Tag Nrho RNA nyob rau hauv thaj tsam ntawm 1pg-1μg, thiab kev sim ntawm tib neeg cov qauv qhia pom qhov kev pom zoo ntawm 1 pg-100 ng, tswj rau tag nrho Ct tus nqi hauv thaj tsam ntawm 15-30 raws li qhov tsim nyog.
3) c.Nws raug nquahu kom siv 20μL lossis 50μL kom ntseeg tau tias siv tau thiab rov tsim dua tshiab ntawm lub hom phiaj gene amplification.
4) d.Thov npaj rau hauv lub rooj ntev zaum ultra-huv thiab siv cov lus qhia tsis muaj nuclease residue thiab cov raj tshuaj tiv thaiv;cov lus qhia nrog lim cartridges raug pom zoo.Tsis txhob kis kab mob sib kis thiab aerosol paug.
2.Reaction program
| Lub voj voog kauj ruam | Temp. | Sijhawm | Lub voj voog |
| Rov qab transcription | 50 ℃a | 6 feeb | 1 |
| Thawj denaturation | 95 ℃ | 5 feeb | 1 |
| Amplification cov tshuaj tiv thaiv | 95 ℃ | 15 sec | 40 |
| 60 ℃b | 30 sec | ||
| Melting nkhaus theem | Instrument Defaults | 1 | |
Nco tseg:
1) a.Qhov ntsuas kub rov qab tuaj yeem xaiv tau ntawm 50-55 ° C raws li kev sim xav tau.Rau cov qauv DNA, cov txheej txheem thim rov qab tuaj yeem raug tshem tawm.
2) ib.Hauv cov xwm txheej tshwj xeeb, qhov kub thiab txias tuaj yeem hloov kho raws li tus nqi primer Tm, 60 ° C raug pom zoo.
Sau ntawv
1. Cov khoom no yog siv rau kev tshawb fawb nkaus xwb.
2. Thov ua haujlwm nrog cov tsho hauv tsev kuaj mob thiab cov hnab looj tes pov tseg, kom koj muaj kev nyab xeeb.
