Ngwa Direct RT-qPCR
Nkọwa ngwa:
◮ Kit Direct RT-qPCR, na-eji Foreasy Reverse Transcriptase na Foreasy HS Taq DNA Polymerase na-etolite site na Foregene, yana ihe nchekwa mmeghachi omume pụrụ iche, na-eme ngwa a na nguzogide siri ike na ndakọrịta, yana ọ nwere ike nwalee mmeghachi omume ozugbo iji sistemụ Foregene Lysis dị ka ndebiri.Ngwa a nwere ike itinye maka ngwa nchọpụta COVID-19 nucleic acid yana mmepe ngwa nchọpụta nje nje ndị ọzọ.
Nkọwa
Ngwa RT-qPCR Direct na-enye ihe nchekwa RT-qPCR chịkọtara iche yana enzyme dị elu nke ọmangwakọta (Taq, M-MLV, RI), na-eme ka ọ dị mma maka ịmepụta ngwa azụ azụ na usoro.mmezi, n'ime mfe nkwenye.
Nkọwapụta
500T, 50,000T
Ngwa ngwa
| Ngwa ọdịnaya (Sistemụ mmeghachi omume 20 μl) | IM-05111-01 | IM-05112-01 |
| 500 T | 50,000 T | |
| Ngwakọta Enzyme Direct | 500 μl×1 | 50 ml×1 |
| 2 × Direct RT-qPCR ihe nchekwa | 1 ml×5 | 500 ml×1 |
| Ntuziaka | 1 | 1 |
Nzọụkwụ arụ ọrụ
Tupu nkwadebe reagent, a ga-agbaze reagents niile dị na ngwa ahụ na ụlọ okpomọkụ ma gwakọta ya nke ọma.
A: Kwadebe of ndebiri na reagent
1. Kwadebe ndebiri RNA a kwadebere (A na-atụ aro ka iji Foregene Total RNA Isolation Kit series kit wepụ na sachaa RNA template) ma ọ bụ ngwaahịa cracking (A na-atụ aro ka iji usoro Foregene Lysis), primers kpọmkwem (10 μM) na ihe ndị ọzọ dị mkpa, ngwá ọrụ.
2. Tinye ihe nchekwa 2 × Direct RT-qPCR, RNase-Free ddH2O na 20 × ROX Reference Dye (ọ bụrụ na ọ's mkpa) n'ime igbe nwere ice, na-eme ka ndị a gbazee n'ụzọ nkịtị, ma gwakọta ya nwayọ.
B: Kwadebe of RT-qPCR usoro
Tebụl 1 : Kwadebe of RT-qPCR susoro
| Akụkụ | Olu | Ntinye uche ikpeazụ |
| 2 × Direct RT-qPCR ihe nchekwa | 10 μl | 1× |
| Ngwakọta Enzyme Direct | 1μL | |
| Ntugharị n'ihu (10 μM) | 0.8 μl | 50-900nM |
| Agbaghachi azụ (10 μM) | 0.8 μl | 50-900nM |
| Nyocha (4 μM) | 1μL | 200nM |
| Template (RNA ma ọ bụ Lysate) | X μL | |
| 20×ROX ntụaka agbaji | - | 1* |
| RNase-FreeddH2O | (6.4-X) μL | |
| Mkpokọta olu | 20μL |
C: Tọọ nke RT-qPCR mmeghachi omume mmemme
Tebụl 2 : RT-qPCR reaction mmemme ntọala (maka ihe atụ: abuo-nzọụkwụ usoro)
| Nzọụkwụ | Okpomọkụ | Oge | okirikiri | Ọdịnaya | |
| 1 | 50 ℃ | 15 min | 1* | 1 | Ntugharị azụ azụ |
| 2 | 95 ℃ | 1 min | 1 | Tupu denaturation | |
| 3 | 95 ℃ | 10 sk | 40-45 | Denaturation | |
| 60 ℃ | 30 nkeji 2* | Anneal/Mgbakwunye | |||
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).
