Cell Direct RT qPCR Khoom—Taqman Direct Cell Lysis Cell Npaj Ib-kauj ruam qRT-PCR Cov Khoom Siv Tshawb Fawb
Cov lus piav qhia
Cov khoom no siv cov txheej txheem lysis tsis zoo kom tso tawm RNA sai sai los ntawm cov qauv ntawm tes rau RT-qPCR cov tshuaj tiv thaiv, tshem tawm cov sijhawm siv thiab ua haujlwm RNA purification txheej txheem, thiab tsuas yog 7 feeb kom tau txais cov qauv RNA uas yuav tsum tau, nrog 5 × Direct RT Mix, 2 × Direct qPCR Mix-Taqman tau sai sai thiab ua tau zoo los ntawm lub sijhawm tiag tiag.
5 × Direct RT Mix thiab 2 × Direct qPCR Mix-Taqman muaj zog inhibitor kam rau ua thiab tuaj yeem ua tau zoo thim rov qab thiab tshwj xeeb amplification siv cov lysate ntawm cov qauv los ntsuas raws li tus qauv.Cov reagent muaj Foregene Reverse Transcriptase, Kub D-Taq DNA Polymerase, dNTPs, MgCl2, Reaction Buffer, PCR Optimizer thiab Stabilizer, uas tuaj yeem siv nrog lysis buffer kom sai thiab yooj yim kuaj cov qauv, thiab muaj cov yam ntxwv ntawm siab rhiab heev, tshwj xeeb thiab ruaj khov.
Specifications
200×20μl Rxns, 1000×20μl r xws
Cov khoom siv
| QuickEasyTMCell Direct RT-qPCR Cov Khoom Siv-Taqman | ||||
| Cov khoom siv 20μl qPCR Reaction System | TSO-01021 | TSO-01022 | Nco tseg | |
| 200 T | 1000 T | |||
|
Ib feem I | Buffer CL | 4ml ib | 20 ml ib | Cell Lysis |
| Foregene Protease Plus II | 80l ua | 400l ua | ||
| Buffer ST | 400l ua | 1ml x 2 | ||
|
Ib feem II | DNA Eraser | 80l ua | 400l ua | |
| 5 × Direct RT Mix * | 160l ua | 800l ua | RT | |
| 2 × Direct qPCR Mix-Taqman * | 1ml x 2 | 1.7ml x 6 | qPCR | |
| 20 × ROX Reference Dye | 40l ua | 200 mL | ||
| RNase-Dawb ddH2O | 1.7ml ib | 10 ml | ||
| Phau ntawv qhia | 1 qob | 1 qob | ||
*:Cell Lysis, 5 × Direct RT Mix, 2 × Direct qPCR Mix-Taqman tuaj yeem yuav nyias
Nta & qhov zoo
■Yooj yim thiab siv tau: nrog Cell Direct RT thev naus laus zis, cov qauv RNA tuaj yeem tau txais hauv 7 feeb xwb.
■ Cov qauv xav tau me me, tsawg li 10 lub hlwb tuaj yeem kuaj tau.
■ Kev xa tawm siab: nws tuaj yeem ntes RNA sai sai hauv cov kab mob hauv 384, 96, 24, 12, 6-zoo daim hlau.
■ DNA Eraser tuaj yeem tshem tawm cov genomes sai sai, txo qhov cuam tshuam rau cov txiaj ntsig kev sim tom ntej.
■ Optimized RT thiab qPCR system ua rau ob-kauj ruam RT-PCR thim rov qab tau zoo dua thiab PCR ntau dua, thiab tiv taus ntau dua rau RT-qPCR cov tshuaj tiv thaiv.
Daim ntawv thov Kit
Scope ntawm daim ntawv thov: kab lis kev cai.
- RNA tso tawm los ntawm cov qauv lysis: tsuas yog siv tau rau RT-qPCR qauv ntawm cov khoom siv no.
- Cov khoom siv tuaj yeem siv rau cov hom phiaj hauv qab no: kev tshuaj ntsuam gene qhia, kev txheeb xyuas ntawm siRNA-mediated gene silencing effect, tshuaj ntsuam xyuas tshuaj, thiab lwm yam.
Cia thiab Txee lub neej
Ntu I ntawm cov khoom siv no yuav tsum muab khaws cia ntawm 4℃;Ntu II yuav tsum khaws cia ntawm -20 ℃.
Foregene Protease Plus II yuav tsum khaws cia ntawm 4℃, tsis txhob khov ntawm -20 ℃.
Reagent 2×Direct qPCR Mix-Taqman yuav tsum khaws cia ntawm -20℃hauv qhov tsaus ntuj;yog siv ntau zaus, nws tseem tuaj yeem khaws cia ntawm 4℃ rau lub sijhawm luv luv (siv li ntawm 10 hnub).
Real Time PCR primer tsim qauv
Forward Primer thiab Reverse Primer
Rau Real Time PCR, primer tsim yog ib qho tseem ceeb heev.Primers muaj feem xyuam rau qhov tshwj xeeb thiab kev ua haujlwm ntawm PCR amplification, thiab tuaj yeem tsim nrog siv rau cov hauv qab no:
- Primer ntev: 18-30bp.
- GC cov ntsiab lus: 40-60%.
- Tus nqi Tm: Primer tsim software, xws li Primer 5, tuaj yeem muab tus nqi Tm ntawm primer.Tm qhov tseem ceeb ntawm cov dej ntws thiab cov dej hauv qab yuav tsum nyob ze li qhov ua tau.Cov mis laij lej Tm kuj tseem siv tau: Tm = 4 °C (G + C) + 2 ° C (A + T).Thaum ua PCR, qhov kub thiab txias hauv qab tus nqi primer Tm ntawm 5 ° C feem ntau yog xaiv raws li qhov kub thiab txias (qhov sib piv ntawm qhov kub ntawm qhov annealing tuaj yeem nce qhov tshwj xeeb ntawm cov tshuaj tiv thaiv PCR).
- Primers thiab PCR khoom:
- Tsim primer PCR amplification khoom ntev yog nyiam dua 100-150bp.
- Tsim primers nyob rau hauv lub thib ob structural cheeb tsam ntawm lub template yuav tsum tau zam kom ntau li ntau tau.
- Zam kev tsim ntawm 2 lossis ntau qhov sib ntxiv ntawm lub hauv paus ntawm 3' kawg ntawm cov dej ntws thiab cov dej hauv qab.
- Primer 3′ davhlau ya nyob twg puag tsis tuaj yeem nrog 3 sib law liag G lossis C.
- Cov primers lawv tus kheej tsis tuaj yeem muaj cov qauv sib xyaw ua ke, txwv tsis pub tus qauv hairpin yuav raug tsim, cuam tshuam rau PCR amplification.
- ATCG yuav tsum tau muab faib raws li qhov sib npaug li qhov ua tau hauv cov txheej txheem primer, thiab 3 'terminal puag yuav tsum raug zam raws li T.
Cov ntawv ntxiv1:c uanws DirectRT-qPCR Kit component ntxiv pob
1.Cell Lysis Solution
| Cell Lysis Solution | |||
| Cov khoom siv (24-zoo lysis system / zoo) | DRT-01011-A1 | DRT-01011-A2 | |
| 100 T | 500 T | ||
| Ib feemKuv | Buffer CL | 20 ml ib | 100 ml |
| Foregene Protease Plus II | 400l ua | 1ml x 2 | |
| Buffer ST | 1ml x 2 | 10 ml | |
| Ib feemII | DNA Eraser | 400l ua | 1ml x 2 |
| RT Mix | |
| Cov khoom siv (20 μl cov tshuaj tiv thaiv kab mob) | DRT-01011-B1 |
| 200 T | |
| 5 × Direct RT Mix | 800l ua |
| RNase-Dawb ddH2O | 1.7ml x 2 |
| qPCR sib | ||
| Cov khoom siv (20 μl cov tshuaj tiv thaiv kab mob) | DRT-01021-C1 | DRT-01021-C2 |
| 200 T | 1000 T | |
| 2 × Direct qPCR Mix-Taqman | 1ml x 2 | 1.7ml x 6 |
| 20 × ROX Siv Dye | 40l ua | 200 mL |
| RNase-Dawb ddH2O | 1.7ml ib | 10 ml |
Phau ntawv qhia:
