Fa'atonu RT-qPCR Kit II
Fa'amatalaga o pusa:
◮Direct RT-qPCR Kit, faʻaaoga Foreasy Reverse Transcriptase ma Foreasy HS Taq DNA Polymerase atiaʻe e Foregene, faʻatasi ai ma faʻalavelave tali faʻapitoa, e faia ai lenei pusa ma malosi malosi ma fetaui, ma e mafai ona suʻeina saʻo le faʻaogaina o le Foregene Lysis system e fai ma faʻataʻitaʻiga.O lenei pusa e mafai ona faʻaoga mo le COVID-19 nucleic acid detection kit ma isi pathogenic bacteria nucleic acid detection kit development kit.
Fa'amatalaga
Direct RT-qPCR Kit e tu'u ese'ese le afifiina o le RT-qPCR pa puipui, maualuga le lelei Foreasy HS Taq DNA Polymerasema Foreasy Reverse Transcriptase(MMLV),fa'afaigofie mo le fa'atupuina o pusa pito i tua ma le fa'aogaina o faiga, i totonu o fa'amaoniga faigofie.
Direct RT-qPCR Kit, faʻaaoga Foreasy Reverse Transcriptase ma Foreasy HS Taq DNA Polymerase atiaʻe e Foregene, faʻatasi ai ma faʻalavelave tali faʻapitoa, e faia ai lenei pusa ma malosi malosi ma fetaui, ma e mafai ona suʻeina saʻo le faʻaogaina o le Foregene Lysis system e fai ma faʻataʻitaʻiga.O lenei pusa e mafai ona faʻaoga mo le COVID-19 nucleic acid detection kit ma isi pathogenic bacteria nucleic acid detection kit development kit.
O lenei pusa e mafai ona avea saʻo ma vaega o le oloa IVD, faʻaoga faigofie ma vave.E manaʻomia naʻo le faʻamaonia faigofie, e aunoa ma le toe faʻaleleia.
Fa'amatalaga
500T, 50,000T
Vaega o pusa
| mea i totonu o pusa (20 μL faiga tali) | IM-05111-02 | IM-05112-02 |
| 500 T | 50,000 T | |
| Foreasy Reverse Transcriptase (200 U/μL) | 50 μL×1 | 1 mL×5 |
| Foreasy HS Taq DNA Polymerase (5 U/μL) | 100 μL×1 | 1 mL×10 |
| 2× Direct RT-qPCRPuipui | 1 mL ×5 | 500 mL ×1 |
| Faatonuga | 1 | 1 |
Fa'agaioiga Laasaga
A: Sauni le faʻataʻitaʻiga ma le reagent
1. Saunia RNA template saunia (E fautuaina e faaaoga Foregene Total RNA Isolation Kit series kit e aveese ma faamama RNA template) poʻo le faʻataʻitaʻiga oloa taʻe (E fautuaina le faʻaogaina o le Foregene Lysis system), mea faʻapitoa (10 μM) ma isi mea faʻaaogaina talafeagai, meafaigaluega.
2. Tuu le 2× Direct RT-qPCR Buffer, RNase-Free ddH2O ma le 20× ROX Reference Dye (pe a manaʻomia) i totonu o le atigipusa ma le aisa, faʻafefeteina nei mea, ma faʻafefiloi malie.
B: Saunia o le RT-qPCR system
Maua le afa o le voluma o le tali pa'u o tali faiga (mo se fa'ata'ita'iga, afai o le voluma o le faiga e 20 μL, e mana'omia ona maua le 10 μL 2× Direct RT-qPCR Buffer). Motutusa le aofaiga ole enzyme, matou te fautua atu lena meaM-MLV:10-30 U/20 μL faiga tali,TaqDNA polymerase：1-2 U/20 μL faiga talio le matou fautuaga lea, e tatau ona e suʻeina ma fetuunai), fa'aopoopo RNA fa'ata'ita'iga, fa'amatalaga fa'apitoa ma su'esu'ega, ma fa'aopoopo le RNase-Free ddH2O i le 20 μL.Ole tapenaga fa'apitoa ole faiga ole tali ole RT-qPCR e mafai ona fa'asino ile laulau 1 o lo'o i lalo.
Laulau 1: Sauni le RT-qPCR system
| Vaega | Volume | Fa'atonuga mulimuli |
| 2× Direct RT-qPCR Pusa | 10 μL | 1× |
| Foreasy Reverse Transcriptase(MMLV) | 10-30 U | |
| Foreasy HS Taq DNA Polymerase | 1-2 U | |
| Fa'amuamua (10 μM) | 0.8 μL | 50-900nM |
| Fa'ata'ita'i Fa'asaga (10 μM) | 0.8 μL | 50-900nM |
| Su'ega(4 μM) | 1 μL | 200nM |
| Fa'ata'ita'iga(RNA po'o le Lysate) | X μL | |
| 20× ROX Fa'ailoga Fa'ailoga | - | 1* |
| RNase-FreeddH2O | (6.4-X) μL | |
| Aofa'i Volume | 20 μL |
Fa'aaliga: Fa'amatalaga Fa'asaga i luma ma le Fa'ailoga Fa'asolo o ni fa'amatalaga fa'apitoa o le kenera fa'atatau.O le faiga o le qPCR e mafai ona fetuunai e tusa ai ma faʻataʻitaʻiga faʻataʻitaʻiga ma PCR faʻataʻitaʻiga.Matou te fautuaina le 400nM mo le faʻatonuga mulimuli o le tele o primers.Fetuuna'i le voluma o fa'amatalaga fa'apitoa ma su'esu'ega e tusa ai ma le fa'atonuga saunia ma le fa'atonuga fa'ai'u fa'amolemole, fa'amolemole.
1*: Filifili le fa'atonuga fa'ai'u talafeagai ole ROX Reference Dye e tusa ai ma meafaigāluega PCR quantitative eseese.Ole maualuga ole ROX Reference Dye concentration o meafaigāluega PCR fa'atusatusa masani o lo'o fa'aalia ile laulau o lo'o mulimuli mai:
| Meafaigāluega PCR aofa'i | Fa'ai'uga fa'ai'uga ole ROX Reference Dye |
| ABI PRISM 7000/7300/7700/7900HT/Laasaga Muamua,ma isi. | 1× (mo se faʻataʻitaʻiga, faʻaopoopo le 1μl 20×ROX Reference Dye i le 20 μL reaction system) |
| ABI 7500, 7500 Anapogi, Stratagene Mx3000P, Mx3005P ma Mx4000, ma isi. | 0.5 × (mo se faʻataʻitaʻiga, faʻaopoopo le 0.5μl 20 × ROX Faʻailoga Faʻailoga i le 20 μL faiga tali) |
| Roche PCR meafaigaluega, Bio-Rad PCR meafaigaluega, Eppendorf quantitative PCR meafaigaluega, ma isi. | A aunoa ma le ROX Reference Dye |
Teuina ma le ola fa'asao
E tatau ona teuina le pusa ile -20 ± 5℃.E tatau ona teuina vave i totonu -20 ℃ pusaaisa thermostatic.Ole taimi fa'agata e 2 tausaga pe a teuina ile tulaga talafeagai.
