Multiplex One Step RT-qPCR Premix
Kufotokozera
Nambala ya mphaka: HCR5141A
Multiplex One Step RT-qPCR Premix ndi multiplex quantitative PCR Kit yozikidwa pa RNA monga template.Poyesera, zolembera zosinthika ndi kuchuluka kwa PCR zimachitidwa mu chubu chofananira, kufewetsa ntchito yoyesera ndikuchepetsa chiopsezo cha kuipitsidwa.Mapangidwe apadera a buffer ndi enzyme mix angagwiritsidwe ntchito mu gawo limodzi la lyophilized system.Chidachi chimagwiritsa ntchito Reverse Transcriptase yosamva kutentha popanga bwino cDNA ya chingwe choyamba pogwiritsa ntchito hotstart Taq DNA Polymerase pakukulitsa kuchuluka.Imakhala ndi buffer yowoneka bwino, kusakanikirana kwa ma enzyme ndi zina, ndi zinthu zomwe zimalepheretsa kukulitsa kwa PCR kosatchulika ndikuwonjezera kukulitsa kwa machitidwe angapo a qPCR zidawonjezedwa, kuthandizira kukulitsa kachulukidwe ka fluorescence ndikuwonetsetsa kuti zoyambira zimakulirakulira.
Zigawo
| Dzina |
| 1. Lyo-Buffer |
| 2. Kusakaniza kwa Lyo-Enzyme |
| 3. Mtetezi wa Lyo |
Njira yamayendedweion
A: Lyo-buffer ndi chitetezo: -25~-15 ℃, alumali moyo ndi 1 chaka.
B: Kusakaniza kwa Lyo-enzyme, 2-8 ℃, moyo wa alumali ndi miyezi 6.
Malangizo ogwiritsira ntchito
1. Dongosolo Lochita (Tengani chitsanzo cha 25μL)
| Zigawo | Kuchuluka (μL) | Kukhazikika Kwambiri |
| Lyo-Buffer | 6 | 1* |
| Lyo-Enzyme Mix | 1 | - |
| Lyo-Protectant | 8 | - |
| Primer Mix (10μM) | 1 | 0.1-1uM |
| Probe Mix (10μM) | 0.5 | 0.05-0.5uM |
| Chithunzi cha RNA | 5 | - |
| DEPC H2O | Mpaka 25 | - |
2. Kukometsedwa Pang'onopang'ono Protocol
1) Standard Cycling Protocol
| Zochita siteji | Kutentha | Nthawi | Kuzungulira | |
| 1 | Sinthani zomasulira | 50°Ca | 10 min | 1 |
| 2 | Denaturation koyamba | 95°C | 5 min | 1 |
| 3 | Amplification reaction | 95°C | 15mphindi | 45 zozungulira |
| 60°Cb | 30mphindic |
2) Fast Cycling Protocol
|
| Zochita siteji | Kutentha | Nthawi | Kuzungulira |
| 1 | Sinthani zomasulira | 50°Ca | 2 min | 1 |
| 2 | Denaturation koyamba | 95°C | 2mphindi | 1 |
| 3 | Amplification reaction | 95°C | 1mphindi |
45 zozungulira |
| 60°Cb | 13mphindic |
Zindikirani:
a) M'mbuyo zolemba: Kutentha kumatha kusankha 42 ° C kapena 50 ° C kwa mphindi 10-15.
b) Amplification reaction: Kutentha kumasinthidwa molingana ndi mtengo wa Tm wa zoyambira zopangidwa.
c)Fluorescence chizindikiro kupeza: Chonde ikani njira yoyeserera molingana ndi zofunikira zachida buku.
Zambiri Zaukadaulo / Zofotokozera
| Hot Start | Yoyambira yotentha yomangidwa |
| Njira yodziwira | Kuzindikira koyambirira |
| Njira ya PCR | Gawo limodzi RT-qPCR |
| Mtundu wa zitsanzo | RNA |
Zolemba
1. Izi ndizogwiritsidwa ntchito pofufuza kokha.
2. Chonde valani PPE yofunikira, malaya a labu ndi magolovesi, kuti mutsimikizire thanzi lanu ndi chitetezo!
