I-ForeDirect RT-qPCR Kit
Inkcazelo yeKit:
◮Kwi-RNA yokukhulisa ixesha lokwenyani ngqo ukusuka kwingqokelela ye-swab ngaphandle kweenkqubo zokucoca i-RNA.Le khithi igqiba imijikelo ye-qRT-PCR kwiyure enye.Umxube we-enzyme ngumxube ophuculweyo we-Reverse Transcriptase, i-Hot-Start Taq DNA polymerase, i-RNase inhibitor.I-Reaction Buffer iqulathe onke amacandelo afunekayo, kubandakanywa amacandelo esikhuseli esiphuculweyo, uMg2+, i-dUTP, kunye ne-dNTPs.
Inkcazo
I-ForeDirect RT-qPCR Kit yenzelwe i-RNA yokukhulisa ixesha lokwenyani ngqo ukusuka kwingqokelela yeswab ngaphandle kweenkqubo zokucoca i-RNA.Le khithi igqiba imijikelo ye-qRT-PCR kwiyure enye.Umxube we-enzyme ngumxube ophuculweyo we-Reverse Transcriptase, i-Hot-Start Taq DNA polymerase, i-RNase inhibitor.I-Reaction Buffer iqulathe onke amacandelo afunekayo, kubandakanywa amacandelo esikhuseli esiphuculweyo, uMg2+, i-dUTP, kunye ne-dNTPs.
Iinkcukacha
100T
Amacandelo ekhithi
| I-agent yokukhululwa kwe-Nucleic acid |
| Umkhuseli we-RNA |
| 2× RT-qPCR Buffer |
| Umxube we-Enzyme(Taq&M-MLV) |
| Imiyalelo |
Isicelo sePCR
1. Nje ukuba iityhubhu ze-PCR zokusabela zifakwe ngokufutshane, zibeke kwitanki yesampula yesixhobo sokukhulisa.
2. Iprotocol ye-Thermal
| Inyathelo | Ubushushu | Ixesha | Imijikelo | |
| 1 | Reverse transcription | 50℃ | 15 imiz | 1 |
| 2 | I-denaturation yangaphambili | 95℃ | 1 min | 1 |
| 3 | I-Denaturation | 95℃ | Imizuzwana eyi-10 | 42 |
| 4 | Isihlomelo/Ukwandiswa | 60℃ | Imizuzwana engama-30 | |
Phawula:Isignali ye-Fluorescent yafunyanwa ngokukhawuleza emva kwesinyathelo sokwandiswa komjikelezo ngamnye.
3. Emva kokucwangcisa, gcina ifayile kwaye usebenzise inkqubo yokusabela.
Amanyathelo okusebenza
Ngaphambi kokulungiswa kwe-reagent, zonke ii-reagents kwikiti kufuneka zincibilike kwiqondo lokushisa elisezantsi kwaye zixutywe ngobumnene.
Ukulungiswa kwe-reagent
1. Lungiselela iNucleic Acid Release Mix
| Icandelo | Umthamo weSampuli nganye okanye uLawulo |
| I-agent yokukhululwa kwe-Nucleic acid | 5μL |
| Umkhuseli we-RNA | 0.5 μL |
2. Lungisa iRection Mix
| Icandelo | Umthamo weSampuli nganye okanye uLawulo |
| 2× RT-qPCR Buffer | 15 μL |
| Umxube we-Enzyme(Taq&M-MLV) | 1.5 μL |
| Iiprimers kunye neProbes | 1μL |
Ugcino kunye nobomi beShelf
-Ikhithi kufuneka igcinwe kwi -20± 5℃.Ixesha lokuqinisekisa ziinyanga ezili-12.
-Kuphephe ukuphindaphinda imijikelo ye-freeze-thaw (<5 imijikelo).
