(96-zoo) QuickEasy Cell Direct RT-qPCR Cov Khoom Siv - SYBR Green I
Cov lus piav qhia
Cov(96-zoo) QuickEasyTM Cell Direct RT-qPCR cov khoom siv-SYBR Green Imuabib qho tshwj xeeb lysis buffer systemto sai sai tso RNAlos ntawm kab lis kev caicov qauv rau RT-qPCR cov tshuaj tiv thaiv, tshem tawm lub sijhawm siv thiab ua haujlwm hnyavRNA purification txheej txheem, thiab tsuas yog siv sijhawm 7 feeb kom tau txais cov qauv RNA xav tau.Cov5 × Direct RT Mixthiab2 × Direct qPCR Mix-SYBRmuab rau hauv lub thawv tuaj yeem sai thiabtau txais txiaj ntsig ntawm lub sijhawm tiag tiagCov txiaj ntsig PCR.
5 × Direct RT Mix thiab 2 × Direct qPCR Mix-SYBR muaj zog inhibitor kam rau ua, thiab tuaj yeem siv cov lysate ntawm cov qauv los kuaj ua tus qauv rau kev rov qab ua haujlwm tau zoo thiab kev ua kom muaj zog.Cov reagent muaj Foregene tshwj xeeb Reverse Transcriptase nrog siab affinity rau RNA, nrog rau kub D-Taq DNA Polymerase, dNTPs, MgCl2 , cov tshuaj tiv thaiv tsis zoo, PCR optimizer thiab stabilizer, thiab tuaj yeem siv ua ke nrog cov lysis tsis tuaj yeem kuaj xyuas cov qauv sai, yooj yim thiab raug, thiab nws muaj cov yam ntxwv ntawm siab rhiab heev, muaj zog tshwj xeeb thiab ruaj khov zoo.
Cov khoom siv yog tsom rau micro-system lysis ntawm 96-zoo kab lis kev cai hlwb, thiab muaj kev sib raug zoo thiab sib xws;Cov khoom siv khoom siv muab 96 lysis cov tshuaj tiv thaiv, 96 qhov kev hloov pauv rov qab thiab 96 × 2 qPCR cov tshuaj tiv thaiv, uas tuaj yeem ua tau raws li 96-zoo hlwb phaj rau kev siv ib zaug, zam cov pa phem los ntawm kev rov qhib, khov thiab thawing ntawm reagents thiab degradation ntawm reagent kev ua tau zoo.
Cov khoom siv
| Cov khoom siv muaj pes tsawg leeg ( 50 μL lysis system/20l mivRT tshuaj tiv thaiv kab mob / 20μL qPCR cov tshuaj tiv thaiv) | DRT-03011 ib | Lus ceeb toom | |
| 96t ua | |||
| Ib feemI | Tsis pubCL | 5 ml ua | CellLysis |
| ForegeneProtease Plus II | 100 μL | ||
| Tsis pubST | 500 μL | ||
| Ib feem II | DNAlwv | 100 μL | |
| 5 × Direct RT Mix | 400 μL | RT | |
| 2 × Direct qPCR Mix-SYBR | 1 m ibL × 2 | qPCR | |
| 50 × ROX Reference Dye | 400µL | ||
| RNase- DawbddH ua2 O | 1.7ml ib |
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| Mtxhua xyoo | 1 ib daim | 1 pab | |
*: Lub lysis reagent DNA Eraser muaj nyob rau hauv Tshooj II ntawm cov khoom siv;Cell Lysis, RT, thiab qPCR Cheebtsam tuaj yeem muas 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:
