o Masini Ma'a Saina
O lenei pusa o loʻo faʻaaogaina se faiga faʻapipiʻi lysis faʻapitoa e mafai ona vave faʻasaʻo le RNA mai faʻataʻitaʻiga faʻaleaganuʻu mo le RT-qPCR tali, ma faʻaumatia ai le taimi faʻaalu ma le galue RNA faʻamamaina.E mafai ona maua le mamanu RNA ile na'o le 7 minute.O le 5×Direct RT Mix ma le 2×Direct qPCR Mix-SYBR reagents e saunia e le pusa e mafai ona vave ma lelei ona maua i'uga o le PCR fa'atatau ile taimi moni.
5 × Direct RT Mix ma 2 × Direct qPCR Mix-SYBR o loʻo i ai le faʻapalepale malosi o le inhibitor, ma o le lysate o faʻataʻitaʻiga e mafai ona faʻaaogaina e fai ma faʻataʻitaʻiga mo RT-qPCR saʻo.O lenei pusa o loʻo i ai le RNA maualuga-affinity Foregene reverse transcriptase, ma Hot D-Taq DNA polymerase, dNTPs, MgCl2, tali fa'afefete, PCR optimizer ma fa'amautu.
200×20μl Rxns, 1000×20μl Rxns
Vaega I | Puipui CL |
Foregene Protease Plus II | |
Puipui ST | |
Vaega II | DNA tape |
5× Direct RT Mix | |
2× Direct qPCR Mix-SYBR | |
50× ROX Fa'ailoga Fa'ailoga | |
RNase-Free ddH2O | |
Faatonuga |
■ Faigofie ma aoga : faʻatasi ai ma le Cell Direct RT tekinolosi, RNA faʻataʻitaʻiga e mafai ona maua i totonu o le 7 minute.
■ O le faʻataʻitaʻiga manaʻoga e laʻititi, e pei o le 10 sela e mafai ona suʻeina.
■ Tulaga maualuga: e mafai ona vave iloa le RNA i sela o loʻo faʻatupuina ile 384, 96, 24, 12, 6-well plates.
■ DNA Eraser e mafai ona vave aveese genomes tatala, faʻaitiitia le aʻafiaga i faʻataʻitaʻiga mulimuli ane.
■ Optimized RT ma qPCR system e sili atu ona lelei le fa'aliliuina o le RT-PCR la'asaga e lua ma sili atu ona patino le PCR, ma sili atu ona fa'asa'o ile fa'ainitaneti tali ole RT-qPCR.
Avanoa o le faʻaogaina: sela faʻaleaganuʻu.
- RNA fa'amatuu mai ile fa'ata'ita'iga lysis: na'o le fa'ata'ita'iga RT-qPCR o lenei pusa.
- E mafai ona fa'aoga le pusa mo fa'amoemoega nei: su'esu'ega o fa'amatalaga o ga'o, fa'amaoniga o le siRNA-mediated gene silencing effect, su'esu'ega o vaila'au, ma isi.
Vaega I o lenei pusa e tatau ona teuina i le 4 ℃;Vaega II e tatau ona teuina ile -20℃.
Foregene Protease Plus II e tatau ona teuina i le 4 ℃, aua le fa'aisa ile -20 ℃.
Reagent 2×Direct qPCR Mix-SYBR e tatau ona teuina i le -20 ℃ i le pogisa;pe a fa'aaoga soo, e mafai foi ona teuina i le 4 ℃ mo le teuina mo taimi pupuu (fa'aoga i totonu ole 10 aso).
E tusa ai ma ou manaʻoga moni, filifili le ata sili ona talafeagai ma fuafuaga faʻatulagaina