ForeDirect RT-qPCR Kit
Fa'amatalaga o pusa:
◮Mo le RNA tuusaʻo faʻateleina taimi moni mai le aoina o swab e aunoa ma le faʻamamaina o le RNA.O lenei pusa e fa'auma le qRT-PCR i totonu ole itula e tasi.Ole fa'afefiloi ole enzyme ole fa'afefiloi lelei ole Reverse Transcriptase, Hot-Start Taq DNA polymerase, RNase inhibitor.O le Reaction Buffer o loʻo i ai vaega uma e manaʻomia, e aofia ai vaega faʻapipiʻi lelei, Mg2+, dUTP, ma dNTPs.
Fa'amatalaga
ForeDirect RT-qPCR Kit ua mamanuina mo le RNA tuusaʻo faʻateleina taimi moni mai le aoina o swab e aunoa ma le faʻamamaina o le RNA.O lenei pusa e fa'auma le qRT-PCR i totonu ole itula e tasi.Ole fa'afefiloi ole enzyme ole fa'afefiloi lelei ole Reverse Transcriptase, Hot-Start Taq DNA polymerase, RNase inhibitor.O le Reaction Buffer o loʻo i ai vaega uma e manaʻomia, e aofia ai vaega faʻapipiʻi lelei, Mg2+, dUTP, ma dNTPs.
Fa'amatalaga
100T
Vaega o pusa
| Nucleic acid fa'asa'olotoga |
| RNA puipuia |
| 2× RT-qPCR Pusa |
| Fa'afefiloi ole Enzyme(Taq&M-MLV) |
| Faatonuga |
Talosaga PCR
1. A uma loa ona fa'a'ese'ese pupuu faagaau tali PCR, tu'u i totonu o le tane fa'ata'ita'i o meafaigāluega fa'aopoopo.
2. Polokalama vevela
| Laa | vevela | Taimi | Ta'amilosaga | |
| 1 | Fa'aliliuga fa'aliliuga | 50℃ | 15 minu | 1 |
| 2 | mua'i denaturation | 95℃ | 1 minu | 1 |
| 3 | Denaturation | 95℃ | 10 sekone | 42 |
| 4 | Anneal/Fa'aopoopoga | 60℃ | 30 sekone | |
Fa'aaliga:Na iloa vave le fa'ailoga o le fluorescent pe a mae'a le la'asaga fa'aopoopo o ta'amilosaga ta'itasi.
3. A maeʻa ona faʻatulagaina, faʻasaoina le faila ma faʻatautaia le polokalama tali.
Fa'agaioiga Laasaga
Aʻo leʻi saunia faʻamalosi, e tatau ona faʻafefe uma mea faʻapipiʻi i totonu o le pusa i le vevela o le potu ma faʻafefiloi malie.
Sauniuniga reagent
1. Saunia le Nucleic Acid Release Mix
| Vaega | Volume i le Fa'ata'ita'iga po'o le Pulea |
| Nucleic acid fa'asa'olotoga | 5μL |
| RNA puipuia | 0.5 μL |
2. Saunia le Reaction Mix
| Vaega | Volume i le Fa'ata'ita'iga po'o le Pulea |
| 2× RT-qPCR Pusa | 15 μL |
| Fa'afefiloi ole Enzyme(Taq&M-MLV) | 1.5 μL |
| Primers ma Su'esu'ega | 1μL |
Teuina ma le ola fa'asao
-O le pusa e tatau ona teuina ile -20± 5℃.Ole vaitaimi ole aoga ole 12 masina.
- Aloese mai ta'amilosaga fa'a'aisa fa'aisa (<5 ta'amilosaga).
