Multiplex One Mohato RT-qPCR Premix
Tlhaloso
Nomoro ea katse: HCR5141A
Multiplex One Step RT-qPCR Premix ke multiplex quantitative PCR Kit e thehiloeng ho RNA e le template.Tekong, reverse transcript le quantitative PCR li etsoa ka tube e ts'oanang ea karabelo, ho nolofatsa ts'ebetso ea liteko le ho fokotsa kotsi ea tšoaetso.Moralo o ikhethileng oa motsoako oa buffer le enzyme o ka sebelisoa mohatong o le mong oa lyophilized system.Setsi sena se sebelisa Reverse Transcriptase e thibelang mocheso bakeng sa ho kopanya hantle cDNA ea pele e sebelisa hotstart Taq DNA Polymerase bakeng sa ho holisa bongata.E na le buffer e ntlafalitsoeng ea karabelo, motsoako oa li-enzyme jj., le lintlha tse thibelang katoloso e sa tobang ea PCR le ho ntlafatsa katleho ea karabelo e mengata ea qPCR e kenyellelitsoe, e nolofalletsang ho holisa bongata ba fluorescence ha ho ntse ho netefatsa katleho ea ho holisa li-primers.
Likaroloana
| Lebitso |
| 1. Lyo-Buffer |
| 2. Motsoako oa Lyo-Enzyme |
| 3. Mosireletsi wa Lyo |
Sepalangoangion
A: Lyo-buffer and protectant: -25~-15℃, shelf bophelo ke 1 selemo.
B: Motsoako oa Lyo-enzyme, 2-8 ℃, bophelo ba sethala ke likhoeli tse 6.
Taelo ea ho sebetsa
1. Rection System (Nka 25μL e le mohlala)
| Likaroloana | Bolumo (μL) | Boikutlo ba ho Qetela |
| Lyo-Buffer | 6 | 1* |
| Motsoako oa Lyo-Enzyme | 1 | - |
| Mosireletsi oa Lyo | 8 | - |
| Primer Mix (10μM) | 1 | 0.1-1uM |
| Probe Mix (10μM) | 0.5 | 0.05-0.5uM |
| Setšoantšo sa RNA | 5 | - |
| DEPC H2O | Ho fihlela ho 25 | - |
2. Optimized Cycling Protocol
1) Protocol e Tloaelehileng ea Libaesekele
| Mokhahlelo oa ho itšoara | Mocheso | Nako | Baesekele | |
| 1 | Phetolelo ea morao | 50°Ca | 10 mets | 1 |
| 2 | Denaturation ea pele | 95°C | 5 mets | 1 |
| 3 | Boitšoaro ba amplification | 95°C | 15sec | 45 potoloho |
| 60°Cb | 30secc |
2) Protocol ea Potlako ea Libaesekele
|
| Mokhahlelo oa ho itšoara | Mocheso | Nako | Baesekele |
| 1 | Phetolelo ea morao | 50°Ca | 2 mets | 1 |
| 2 | Denaturation ea pele | 95°C | 2mts | 1 |
| 3 | Boitšoaro ba amplification | 95°C | 1mts |
45 potoloho |
| 60°Cb | 13secc |
Hlokomela:
a) Kgutla sengoliloeng: Mocheso o ka khetha 42°C kapa 50°C ka metsotso e 10-15.
b) Boitšoaro ba amplification: Mocheso o lokisoa ho ea ka boleng ba Tm ba li-primers tse entsoeng.
c)Fluorescence sesupo ho fumana: Ka kopo, beha mokhoa oa liteko ho latela litlhoko tsabukana ea lisebelisoa.
Lintlha tsa Theknoloji / Litlhaloso
| Hot Qala | Ho qala ho chesa |
| Mokhoa oa ho lemoha | Ho fumanoa ka lekhetlo la pele |
| Mokhoa oa ho sebelisa PCR | Mohato o le mong RT-qPCR |
| Mofuta oa mohlala | RNA |
Lintlha
1. Sehlahisoa sena ke sa tšebeliso ea lipatlisiso feela.
2. Ka kopo, roala PPE e hlokahalang, seaparo se joalo sa lab le liatlana, ho netefatsa bophelo bo botle le polokeho ea hau!
