Multiplex Tasi Laasaga RT-qPCR Premix
Fa'amatalaga
Numera pusi: HCR5141A
Multiplex One Step RT-qPCR Premix ose PCR Kit e fa'atatau ile RNA e fai ma fa'ata'ita'iga.I le fa'ata'ita'iga, o le fa'aliliuina o le fa'aliliuga ma le PCR fa'atusatusa e fa'atino i totonu o le fa'aputuga tali tutusa, fa'afaigofie le fa'ata'ita'iga fa'ata'ita'iga ma fa'aitiitia le lamatiaga o le fa'aleagaina.O le mamanu tulaga ese o le paluga ma le enzyme mix e mafai ona faʻaaogaina i le tasi-laasaga lyophilized system.O lo'o fa'aogaina e le pusa le Reverse Transcriptase e fa'asa'o vevela mo le fa'aogaina lelei o le strand cDNA muamua e fa'aaoga ai le Hotstart Taq DNA Polymerase mo le fa'ateleina fa'atele.O loʻo i ai le faʻaleleia atili o tali, faʻafefiloi enzymes ma isi mea, ma mea taua e taofia ai le faʻalauteleina o le PCR e le faʻapitoa ma faʻaleleia le faʻaogaina o le tele o tali qPCR na faʻaopoopoina, faʻatagaina le tele o le fluorescence quantitative amplification aʻo faʻamautinoa le faʻamalosia lelei o primers.
Vaega
| Igoa |
| 1. Lyo-Buffer |
| 2. Faafefiloi Lyo-Enzyme |
| 3. Lyo puipuia |
Tulaga o felauaigaion
A: Lyo-buffer ma le puipuiga: -25 ~ -15 ℃, ola fata e 1 tausaga.
B: fa'afefiloi Lyo-enzyme, 2-8 ℃, ola fa'aola e 6 masina.
Faatonuga mo le faagaoioiga
1. Reaction System (Ave 25μL e fai ma faʻataʻitaʻiga)
| Vaega | Volume (μL) | Fa'ai'uga Fa'amau |
| Lyo-Buffer | 6 | 1* |
| Faafefiloi Lyo-enzyme | 1 | - |
| Lyo-Puipui | 8 | - |
| Fa'afefiloi Muamua (10μM) | 1 | 0.1- 1uM |
| Fa'afefiloi Su'esu'e (10μM) | 0.5 | 0.05-0.5uM |
| RNA Fa'ata'ita'iga | 5 | - |
| DEPC H2O | E oʻo atu i le 25 | - |
2. Fa'akomepiuta Fa'ata'amilosaga Lelei
1) Polokalama Fa'ata'avalevale Fa'ata'avalevale
| Laasaga tali | vevela | Taimi | Ta'amilosaga | |
| 1 | Fa'aliliuga fa'aliliuga | 50°Ca | 10min | 1 |
| 2 | Uluai denaturation | 95°C | 5min | 1 |
| 3 | Tali fa'atuputeleina | 95°C | 15sec | 45 taamilosaga |
| 60°Cb | 30secc |
2) Polokalama Ti'eti'e uila vave
|
| Laasaga tali | vevela | Taimi | Ta'amilosaga |
| 1 | Fa'aliliuga fa'aliliuga | 50°Ca | 2min | 1 |
| 2 | Uluai denaturation | 95°C | 2sec | 1 |
| 3 | Tali fa'atuputeleina | 95°C | 1sec |
45 taamilosaga |
| 60°Cb | 13secc |
Fa'aaliga:
a) Fa'afo'i fa'aliliuga: E mafai e le vevela ona filifili le 42 ° C poʻo le 50 ° C mo le 10-15 minute.
b) Tali fa'atuputeleina: O le vevela e fetuutuunai e tusa ai ma le tau Tm o mea na fuafuaina.
i)Fluorescence faailoilo mauaina: Fa'amolemole seti le faiga fa'ata'ita'i e tusa ai ma mana'oga o letusi lesona meafaigaluega.
Faʻamatalaga Faʻamatalaga/Faʻamatalaga
| Amata vevela | Fa'atosina vevela amata |
| Su'esu'ega auala | Su'esu'ega muamua |
| auala PCR | Tasi le laasaga RT-qPCR |
| Ituaiga fa'ata'ita'iga | RNA |
Fa'amatalaga
1. O lenei oloa e mo na'o su'esu'ega.
2. Fa'amolemole fai le PPE e mana'omia, la'ei fale faigaluega ma totigilima, e fa'amautinoa ai lou soifua maloloina ma le saogalemu！
