Kit ForeDirect RT-qPCR
Nkọwa ngwa:
◮Maka nkwalite RNA ozugbo sitere na mkpokọta swab na-enweghị usoro nchacha RNA.Ngwa a na-emecha okirikiri qRT-PCR n'ime otu elekere.Ngwakọta enzyme bụ ngwakọta kachasị nke Reverse Transcriptase, Hot-Start Taq DNA polymerase, RNase inhibitor.Ihe nchekwa mmeghachi omume nwere ihe niile achọrọ, gụnyere akụrụngwa nchekwa kachasị, Mg2+, dUTP, na dNTPs.
Nkọwa
Emebere Kit ForeDirect RT-qPCR maka nkwalite RNA ozugbo site na mkpokọta swab na-enweghị usoro nchacha RNA.Ngwa a na-emecha okirikiri qRT-PCR n'ime otu elekere.Ngwakọta enzyme bụ ngwakọta kachasị nke Reverse Transcriptase, Hot-Start Taq DNA polymerase, RNase inhibitor.Ihe nchekwa mmeghachi omume nwere ihe niile achọrọ, gụnyere akụrụngwa nchekwa kachasị, Mg2+, dUTP, na dNTPs.
Nkọwapụta
100T
Ngwa ngwa
| Ihe ntọhapụ nke Nucleic acid |
| ihe nchekwa RNA |
| 2 × RT-qPCR ihe nchekwa |
| Ngwakọta Enzyme (Taq&M-MLV) |
| Ntuziaka |
Ngwa PCR
1. Ozugbo PCR mmeghachi omume tubes e nkenke centrifuged, tinye ha na sample tank nke amplification ngwá.
2. Thermal protocol
| Nzọụkwụ | Okpomọkụ | Oge | okirikiri | |
| 1 | Ntugharị azụ azụ | 50 ℃ | Nkeji 15 | 1 |
| 2 | Tupu denaturation | 95 ℃ | 1 min | 1 |
| 3 | Denaturation | 95 ℃ | 10 sekọnd | 42 |
| 4 | Anneal/Mgbakwunye | 60 ℃ | 30 sekọnd | |
Mara:Achọpụtara mgbaàmà fluorescent ozugbo ọ gbatịchara nzọụkwụ nke okirikiri ọ bụla.
3. Mgbe ntọala, chekwaa faịlụ ma mee mmemme mmeghachi omume.
Nzọụkwụ arụ ọrụ
Tupu nkwadebe reagent, a ga-agbaze reagents niile dị na ngwa ahụ na ụlọ okpomọkụ ma gwakọta ya nke ọma.
Nkwadebe reagent
1. Kwadebe Ngwakọta Mwepụta Nucleic Acid
| Akụkụ | Olu kwa Sample ma ọ bụ Njikwa |
| Ihe ntọhapụ nke Nucleic acid | 5μl |
| ihe nchekwa RNA | 0,5 μl |
2. Kwadebe ngwakọta mmeghachi omume
| Akụkụ | Olu kwa Sample ma ọ bụ Njikwa |
| 2 × RT-qPCR ihe nchekwa | 15 μl |
| Ngwakọta Enzyme (Taq&M-MLV) | 1.5 μl |
| Primers na nyocha | 1μl |
Nchekwa na Shelf ndụ
-Ekwesịrị ịchekwa ngwa ahụ na -20± 5℃.Oge nkwado bụ ọnwa iri na abụọ.
-zenarị okirikiri ifriizi ugboro ugboro (<5 okirikiri).
