Direct RT-qPCR Kit
Kufotokozera Kwa Zida:
◮ Direct RT-qPCR Kit, pogwiritsa ntchito Foreasy Reverse Transcriptase ndi Foreasy HS Taq DNA Polymerase yopangidwa ndi Foregene, yokhala ndi buffer yapadera, imapangitsa chida ichi kukhala cholimba komanso chogwirizana, ndipo chimatha kuyesa mwachindunji kugwiritsa ntchito kachitidwe ka Foregene Lysis ngati template.Zidazi zitha kuyika zida zozindikirira za COVID-19 nucleic acid ndi mabakiteriya ena a pathogenic nucleic acid kuzindikira zida.
Kufotokozera
Direct RT-qPCR Kit imapereka RT-qPCR yosungidwa padera komanso enzyme yogwira ntchito kwambiri.osakaniza (Taq, M-MLV, RI), kupanga izo yabwino kwa kulenga zida kumbuyo-mapeto ndi dongosolokusintha, mkati yotsimikizira yosavuta.
Zofotokozera
500T, 50,000T
Zida zamagulu
| Zamkatimu (20 μL reaction system) | IM-05111-01 | IM-05112-01 |
| 500 T | 50,000 T | |
| Direct Enzyme Mix | 500 μL×1 pa | 50 ml pa×1 pa |
| 2 × Direct RT-qPCR Buffer | 1 ml×5 pa | 500 ml×1 pa |
| Malangizo | 1 | 1 |
Njira Zogwirira Ntchito
Pamaso reagent kukonzekera, onse reagents mu zida ayenera thawed firiji ndi kusakaniza mokoma.
A: Konzekerani of template ndi reagent
1. Konzani template ya RNA yokonzekera (Ndibwino kuti mugwiritse ntchito zida za Foregene Total RNA Isolation Kit kuti muchotse ndi kuyeretsa template ya RNA) kapena chitsanzo chophwanyidwa (Ndikulimbikitsidwa kugwiritsa ntchito dongosolo la Foregene Lysis), zoyambira zenizeni (10 μM) ndi zina zowonjezera, chida.
2. Ikani 2 × Direct RT-qPCR Buffer, RNase-Free ddH2O ndi 20× ROX Reference Dye (ngati's zofunika) mu bokosi ndi ayezi, kupanga izi kusungunuka mwachibadwa, ndi kusakaniza mofatsa.
B: Konzekerani of RT-qPCR dongosolo
Table 1 : Konzekerani of RT-qPCR sdongosolo
| Chigawo | Voliyumu | Kukhazikika komaliza |
| 2 × Direct RT-qPCR Buffer | 10 μl pa | 1 × pa |
| Direct Enzyme Mix | 1μL | |
| Woyamba Patsogolo (10 μM) | 0.8 μl | 50-900nM |
| Choyambira Chosinthira (10 μM) | 0.8 μl | 50-900nM |
| Kufufuza (4 μM) | 1μL | 200nM |
| Template (RNA kapena Lysate) | X μL | |
| 20 × ROX Reference Dye | - | 1* |
| RNase-FreeddH2O | (6.4-X) μL | |
| Chiwerengero chonse | 20μL |
C: Khalani ndi RT-qPCR anachita pulogalamu
Table 2 : RT-qPCR anachitaon pulogalamu kukhazikitsa (za chitsanzo: awiri-sitepe njira)
| Khwerero | Kutentha | Nthawi | Zozungulira | Zamkatimu | |
| 1 | 50 ℃ | 15 min | 1* | 1 | Sinthani zomasulira |
| 2 | 95 ℃ | 1 min | 1 | Pre-denaturation | |
| 3 | 95 ℃ | 10 sec | 40-45 | Denaturation | |
| 60 ℃ | 30 mphindi 2 * | Zowonjezera / Zowonjezera | |||
Kusungirako ndi moyo wa alumali
-Zida ziyenera kusungidwa pa -20± 5℃.Nthawi yovomerezeka ndi miyezi 12.
- Pewani kuzungulira kobwerezabwereza kuzizira (<5 mizunguliro).
