Kit ForeDirect RT-qPCR
ʻO ka wehewehe ʻana o ka pahu:
◮No ka hoʻonui maoli ʻana o RNA mai nā hōʻiliʻili swab me ka ʻole o nā kaʻina hoʻomaʻemaʻe RNA.Hoʻopau kēia pahu i nā pōʻai qRT-PCR i loko o hoʻokahi hola.ʻO ka hui ʻana o ka enzyme kahi hui maikaʻi o Reverse Transcriptase, Hot-Start Taq DNA polymerase, RNase inhibitor.Loaʻa i ka Reaction Buffer nā mea i makemake ʻia, me nā ʻāpana buffer optimized, Mg2+, dUTP, a me nā dNTP.
wehewehe
Hoʻolālā ʻia ʻo ForeDirect RT-qPCR Kit no ka hoʻonui ʻana i ka manawa maoli o RNA mai nā hōʻiliʻili swab me ka ʻole o nā kaʻina hoʻomaʻemaʻe RNA.Hoʻopau kēia pahu i nā pōʻai qRT-PCR i loko o hoʻokahi hola.ʻO ka hui ʻana o ka enzyme kahi hui maikaʻi o Reverse Transcriptase, Hot-Start Taq DNA polymerase, RNase inhibitor.Loaʻa i ka Reaction Buffer nā mea i makemake ʻia, me nā ʻāpana buffer optimized, Mg2+, dUTP, a me nā dNTP.
Nā kikoʻī
100T
Nā ʻāpana pahu
| Mea hoʻokuʻu waika nucleic |
| RNA pale |
| 2× RT-qPCR Buffer |
| Huihui Enzyme(Taq&M-MLV) |
| Nā kuhikuhi |
Noi PCR
1. I ka manawa i ho'opōkole 'ia nā paipu hopena PCR, e ho'okomo i loko o ka pahu hāpana o ka mea ho'onui.
2. Kūlana wela
| ʻanuʻu | Mahana | Manawa | Nā pōʻaiapuni | |
| 1 | Hoʻihoʻi i ka palapala | 50 ℃ | 15 min | 1 |
| 2 | Pre-denaturation | 95 ℃ | 1 min | 1 |
| 3 | Denaturation | 95 ℃ | 10 kekona | 42 |
| 4 | Anneal/Hoʻonui | 60 ℃ | 30 kekona | |
Nānā:Ua ʻike ʻia ka hōʻailona Fluorescent ma hope koke o ka hoʻonui ʻia ʻana o kēlā me kēia pōʻai.
3. Ma hope o ka hoʻonohonoho ʻana, mālama i ka faila a holo i ka papahana pane.
Nā ʻanuʻu hana
Ma mua o ka hoʻomākaukau ʻana i ka reagent, pono e hoʻoheheʻe ʻia nā reagents a pau i loko o ka pahu i kahi mahana wela a hui mālie.
Ka hoʻomākaukau ʻana i ka reagent
1. E hoʻomākaukau i ka Nucleic Acid Release Mix
| ʻāpana | Volume no kēlā me kēia laʻana a i ʻole ka mana |
| Mea hoʻokuʻu waika nucleic | 5μL |
| RNA pale | 0.5 μL |
2. E hoʻomākaukau i ka Reaction Mix
| ʻāpana | Volume no kēlā me kēia laʻana a i ʻole ka mana |
| 2× RT-qPCR Buffer | 15 μL |
| Huihui Enzyme(Taq&M-MLV) | 1.5 μL |
| Nā Primers a me nā Probes | 1μL |
Ka mālama ʻana a me ka mālama ʻana
- Pono e mālama ʻia ka pahu ma -20± 5℃.ʻO 12 mahina ka manawa kūpono.
-E hōʻalo i nā pōʻai maloʻo maloʻo (<5 mau pōʻai).
