Ngqo RT-qPCR Kit
Inkcazelo yeKit:
◮ Ngqo RT-qPCR Kit, usebenzisa Foreasy Reverse Transcriptase kunye Foreasy HS Taq DNA Polymerase ephuhliswayo yi Foregene, kunye buffer reaction unique, yenza le khithi ukuxhathisa olomeleleyo kunye nokuhambelana, kwaye inokuvavanya ngokuthe ngqo reaction ukusebenzisa inkqubo Foregene Lysis njenge template.Le khithi inokufaka isicelo sekhithi yokufumanisa i-COVID-19 nucleic acid kunye nezinye iibhaktheriya ze-pathogenic ze-nucleic acid yophuhliso lwekhithi yokufumanisa.
Inkcazo
IKit ye-RT-qPCR ethe ngqo ibonelela ngokwahlukeneyo nge-RT-qPCR buffer kunye ne-enzyme ephezulu esebenzayo.umxube (Taq, M-MLV, RI), iyenza ilungele ukuyilwa kweekhithi zesiphelo kunye nenkqubouhlengahlengiso, ngaphakathi koqinisekiso olulula.
Iinkcukacha
500T, 50,000T
Amacandelo ekhithi
| Imixholo yekhithi (20 μL inkqubo yokusabela) | IM-05111-01 | IM-05112-01 |
| 500 T | 50,000 T | |
| Umxube othe ngqo we-Enzyme | 500 μL×1 | 50 ml×1 |
| 2× Ngqo RT-qPCR Buffer | 1 ml×5 | 500 ml×1 |
| Umyalelo | 1 | 1 |
Amanyathelo okusebenza
Ngaphambi kokulungiswa kwe-reagent, zonke ii-reagents kwikiti kufuneka zincibilike kwiqondo lokushisa elisezantsi kwaye zixutywe ngobumnene.
A: Lungiselela of ithempleyithi kwaye ireagent
1. Lungiselela i-template ye-RNA elungiselelwe (Kucetyiswa ukuba usebenzise i-Foregene Total RNA Isolation Kit series kit ukuze ukhuphe kwaye uhlambulule i-template ye-RNA) okanye isampula yokuqhawula imveliso (Kucetyiswa ukuba kusetyenziswe inkqubo ye-Foregene Lysis), ii-primers ezithile (10 μM) kunye nezinye izinto ezisetyenziswayo ezifanelekileyo, isixhobo.
2. Beka i-2× Direct RT-qPCR Buffer, RNase-Free ddH2O kunye ne-20× ROX Reference Dye (ukuba's ezifunekayo) kwibhokisi enomkhenkce, ukwenza ezi zinyibilike ngokwendalo, kwaye zixutywe ngobunono.
B: Lungiselela of I-RT-qPCR inkqubo
Itheyibhile 1 : Lungiselela of I-RT-qPCR sinkqubo
| Icandelo | Umthamo | Ugxininiso lokugqibela |
| 2× Ngqo RT-qPCR Buffer | 10 μL | 1× |
| Umxube othe ngqo we-Enzyme | 1μL | |
| I-Primer Phambili(10 μM) | 0.8 μL | 50-900nM |
| Reverse Primer(10 μM) | 0.8 μL | 50-900nM |
| Probe(4 μM) | 1μL | 200nM |
| Template(RNA okanye Lysate) | X μL | |
| I-20 × i-ROX ye-Reference Dye | - | 1* |
| RNase-FreeddH2O | (6.4-X) μL | |
| Umthamo opheleleyo | 20μL |
C: Seta i RT-qPCR ukusabela inkqubo
Itheyibhile 2 : I-RT-qPCR sabelaneon inkqubo ulungiselelo (ngenxa umzekelo: Mbini-inyathelo indlela)
| Inyathelo | Ubushushu | Ixesha | Imijikelo | Umxholo | |
| 1 | 50℃ | 15 imiz | 1* | 1 | Reverse transcription |
| 2 | 95℃ | 1 min | 1 | I-denaturation yangaphambili | |
| 3 | 95℃ | 10 imizuzwana | 40-45 | I-Denaturation | |
| 60℃ | 30 imizuzwana 2* | Isihlomelo/Ukwandiswa | |||
Ugcino kunye nobomi beShelf
-Ikhithi kufuneka igcinwe kwi -20± 5℃.Ixesha lokuqinisekisa ziinyanga ezili-12.
-Kuphephe ukuphindaphinda imijikelo ye-freeze-thaw (<5 imijikelo).
