Direct RT-qPCR Kit III
Kit Description:
◮Direct RT-qPCR Cov Khoom Siv, siv Foreasy Reverse Transcriptase thiab Foreasy HS Taq DNA Polymerase tsim los ntawm Foregene, nrog cov tshuaj tiv thaiv tsis zoo, ua rau cov khoom siv no muaj zog tiv thaiv thiab ua tau zoo, thiab nws tuaj yeem kuaj cov tshuaj tiv thaiv ncaj qha siv Foregene Lysis system ua qauv.Cov khoom siv no tuaj yeem thov rau COVID-19 cov khoom siv kuaj pom nucleic acid thiab lwm yam kab mob kab mob nucleic acid nrhiav cov khoom tsim kho.
Kev piav qhia
Direct RT-qPCR Kit muab cais ntim RT-qPCR tsis (nrog Foreasy HS Taq DNA polymerase tov) thiab ua haujlwm zooForeasy Reverse Transcriptase (MMLV),ua kom yooj yim rau kev tsim cov khoom siv rov qab kawg thiab kev hloov kho, hauv kev txheeb xyuas yooj yim.
Direct RT-qPCR Cov Khoom Siv, siv Foreasy Reverse Transcriptase thiab Foreasy HS Taq DNA Polymerase tsim los ntawm Foregene, nrog cov tshuaj tiv thaiv tsis zoo, ua rau cov khoom siv no muaj zog tiv thaiv thiab ua tau zoo, thiab nws tuaj yeem kuaj cov tshuaj tiv thaiv ncaj qha siv Foregene Lysis system ua qauv.Cov khoom siv no tuaj yeem thov rau COVID-19 cov khoom siv kuaj pom nucleic acid thiab lwm yam kab mob kab mob nucleic acid nrhiav cov khoom tsim kho.
Cov khoom siv no tuaj yeem ncaj qha los ntawm cov khoom siv IVD, siv tau yooj yim thiab ceev.Nws tsuas yog xav tau kev txheeb xyuas yooj yim, tsis muaj kev tsim kho dua.
Specifications
500T, 50,000T
Cov khoom siv
| Cov khoom siv (20 μL cov tshuaj tiv thaiv kab mob) | IM-05111-03 ib | IM-05112-03 ib |
| 500 T | 50,000 T | |
| Foreasy REverse Transscriptase(200 UA/μL) | 50l ua× 1 | 1 ml ib× 5 |
| 2 × Direct qPCRMix-Taqman | 1ml x 5 | 500 ml× 1 |
| Kev qhia | 1 | 1 |
Cov kauj ruam ua haujlwm
A: Npaj cov qauv thiab reagent
1. Npaj npaj RNA template (Nws raug nquahu kom siv Foregene Total RNA Isolation Kit series cov khoom siv los rho tawm thiab ua kom huv RNA template) lossis cov khoom lag luam tawg (Nws pom zoo kom siv Foregene Lysis system), cov primers tshwj xeeb (10 μM) thiab lwm yam khoom siv, ntsuas.
2. Muab 2 × Direct RT-qPCR Buffer, RNase-Free ddH2O thiab 20 × ROX Reference Dye (yog tias xav tau) rau hauv lub thawv nrog dej khov, ua rau cov yaj no, thiab sib tov maj mam.
B: Npaj ntawm RT-qPCR system
Tau ib nrab ntawm qhov ntim ntawm cov tshuaj tiv thaiv tsis zoo ntawm cov tshuaj tiv thaiv kab mob (piv txwv li, yog tias qhov ntim ntawm lub kaw lus yog 20 μL, nws yuav tsum tau txais 10 μL 2 × Direct RT-qPCR Buffer). Rautus nqi sib npaug ntawm enzyme, peb xav tiasM-MLV: 10-30 U / 20 μL cov tshuaj tiv thaiv,TaqDNA polymerase:1-2 U / 20 μL cov tshuaj tiv thaivqhov no yog peb qhov kev pom zoo, koj yuav tsum sim thiab kho nws), ntxiv RNA template, tshwj xeeb primers thiab probes, thiab ntxiv RNase-Free ddH2O rau 20 μL.Kev npaj tshwj xeeb ntawm RT-qPCR cov tshuaj tiv thaiv tuaj yeem raug xa mus rau cov lus hauv qab no 1.
Table 1: Npaj ntawm RT-qPCR system
| Cheebtsam | Ntim | Thaum kawg concentration |
| 2 × Direct RT-qPCR Buffer | 10 mL | 1 × |
| Foreasy Reverse Transcriptase (MMLV) | 10-30 Nws | |
| Foreasy HS Taq DNA Polymerase | 1-2 UA | |
| Forward Primer (10 μM) | 0,8l ua | 50-900 nM |
| Rov qab Primer (10 μM) | 0,8l ua | 50-900 nM |
| Kev sojntsuam (4 μM) | 1 mL ib | 200 nM |
| Template (RNA los yog Lysate) | X μL | |
| 20 × ROX Siv Dye | - | 1* |
| RNase-FreeddH2O | (6.4-X) μL | |
| Tag nrho ntim | 20 mL |
Nco tseg: Forward Primer thiab Reverse Primer yog cov primers tshwj xeeb ntawm cov hom phiaj.Lub kaw lus ntawm qPCR tuaj yeem hloov kho raws li kev sim thiab PCR qauv.Peb pom zoo 400nM rau qhov kawg concentration ntawm feem ntau primers.Kho qhov ntim ntawm cov primers tshwj xeeb thiab sojntsuam raws li kev npaj siab thiab qhov pom zoo kawg concentration, thov.
1 *: Xaiv qhov tsim nyog kawg ntawm ROX Reference Dye raws li cov khoom siv PCR sib txawv.Qhov zoo tshaj plaws ROX Reference Dye concentration ntawm cov khoom siv PCR ntau ntau yog qhia hauv cov lus hauv qab no:
| Quantitative PCR instruments | Qhov kawg concentration ntawm ROX Reference Dye |
| ABI PRISM 7000/7300/7700/7900HT / Kauj Ruam Ib,lwm. | 1 × (piv txwv li, ntxiv 1μl 20 × ROX Reference Dye hauv 20 μL cov tshuaj tiv thaiv) |
| ABI 7500, 7500 Fast, Stratagene Mx3000P, Mx3005P thiab Mx4000, thiab lwm yam. | 0.5 × (piv txwv li, ntxiv 0.5μl 20 × ROX Reference Dye hauv 20 μL cov tshuaj tiv thaiv) |
| Roche PCR ntsuas, Bio-Rad PCR ntsuas, Eppendorf quantitative PCR ntsuas, thiab lwm yam. | Tsis muaj ROX Reference Dye |
Cia thiab Txee lub neej
Cov khoom siv yuav tsum khaws cia ntawm -20 ± 5 ℃.Nws yuav tsum tam sim ntawd muab cia rau hauv -20 ℃ thermostatic tub yees.Lub sijhawm siv tau yog 2 xyoos yog tias nws khaws cia rau hauv qhov tsim nyog.
