ForeDirect RT-qPCR Kit
Kufotokozera Kwa Zida:
◮Kwa RNA kukulitsa zenizeni zenizeni kuchokera pazosonkhanitsa za swab popanda njira zoyeretsera RNA.Zidazi zimamaliza kuzungulira kwa qRT-PCR mkati mwa ola limodzi.Kusakaniza kwa enzyme ndi kusakanikirana kokwanira kwa Reverse Transcriptase, Hot-Start Taq DNA polymerase, RNase inhibitor.Reaction Buffer ili ndi zinthu zonse zofunika, kuphatikiza zida zokongoletsedwa bwino, Mg2+, dUTP, ndi dNTPs.
Kufotokozera
ForeDirect RT-qPCR Kit idapangidwa kuti ikhale yokulitsa nthawi yeniyeni ya RNA kuchokera pazosonkhanitsa za swab popanda njira zoyeretsera RNA.Zidazi zimamaliza kuzungulira kwa qRT-PCR mkati mwa ola limodzi.Kusakaniza kwa enzyme ndi kusakanikirana kokwanira kwa Reverse Transcriptase, Hot-Start Taq DNA polymerase, RNase inhibitor.Reaction Buffer ili ndi zinthu zonse zofunika, kuphatikiza zida zokongoletsedwa bwino, Mg2+, dUTP, ndi dNTPs.
Zofotokozera
100T
Zida zamagulu
| Nucleic acid kumasula wothandizira |
| Chitetezo cha RNA |
| 2 × RT-qPCR Buffer |
| Kuphatikiza kwa Enzyme (Taq&M-MLV) |
| Malangizo |
Pulogalamu ya PCR
1. Pamene PCR anachita machubu akhala mwachidule centrifuged, ikani mu thanki chitsanzo cha matalikidwe chida.
2. Thermal protocol
| Khwerero | Kutentha | Nthawi | Zozungulira | |
| 1 | Sinthani zomasulira | 50 ℃ | 15 mins | 1 |
| 2 | Pre-denaturation | 95 ℃ | 1 min | 1 |
| 3 | Denaturation | 95 ℃ | 10 masekondi | 42 |
| 4 | Zowonjezera / Zowonjezera | 60 ℃ | 30 masekondi | |
Zindikirani:Chizindikiro cha fluorescent chinadziwika mwamsanga pambuyo pa sitepe yowonjezera ya kuzungulira kulikonse.
3. Pambuyo poika, kupulumutsa wapamwamba ndi kuthamanga anachita pulogalamu.
Njira Zogwirira Ntchito
Pamaso reagent kukonzekera, onse reagents mu zida ayenera thawed firiji ndi kusakaniza mokoma.
Kukonzekera kwa reagent
1. Konzani Nucleic Acid Release Mix
| Chigawo | Voliyumu pa Sample kapena Control |
| Nucleic acid kumasula wothandizira | 5 μl pa |
| Chitetezo cha RNA | 0.5 μL |
2. Konzani Rection Mix
| Chigawo | Voliyumu pa Sample kapena Control |
| 2 × RT-qPCR Buffer | 15 μl pa |
| Kuphatikiza kwa Enzyme (Taq&M-MLV) | 1.5 μL |
| Zoyamba ndi Probes | 1 μl pa |
Kusungirako ndi moyo wa alumali
-Zida ziyenera kusungidwa pa -20± 5℃.Nthawi yovomerezeka ndi miyezi 12.
- Pewani kuzungulira kobwerezabwereza kuzizira (<5 mizunguliro).
