Multiplex Imwe Nhanho RT-qPCR Premix
Tsanangudzo
Katsi Nhamba: HCR5141A
Multiplex Imwe Nhanho RT-qPCR Premix ndeye multiplex quantitative PCR Kit yakavakirwa paRNA setemplate.Mukuedza, reverse transcription uye quantitative PCR inoitwa mune imwechete reaction chubhu, kurerutsa mashandiro ekuyedza uye kuderedza njodzi yekusvibiswa.Iyo yakasarudzika dhizaini yebuffer uye enzyme musanganiswa inogona kushandiswa mune imwe-nhanho lyophilized system.Iyo kiti inoshandisa kupisa-inodzivirira Reverse Transcriptase kuti ishandiswe zvakanaka yekutanga strand cDNA ichishandisa hotstart Taq DNA Polymerase yekuwedzeredza kuwanda.Iyo ine optimized reaction buffer, enzymes musanganiswa nezvimwe, uye zvinhu zvinonyatso kuvharidzira zvisiri-chaiyo PCR kukwidziridzwa uye kuwedzera kukwidziridzwa kweakawanda qPCR maitiro akawedzerwa, ichigonesa akawanda fluorescence kuwanda kukwidziridzwa uku uchiona kukwidziridzwa kwemaprimers.
Zvikamu
| Zita |
| 1. Lyo-Buffer |
| 2. Lyo-Enzyme Mix |
| 3. Lyo protectant |
Konditi yekufambisaion
A: Lyo-buffer uye inodzivirira: -25~-15 ℃, hupenyu hwesherufu ndeye 1 gore.
B: Lyo-enzyme musanganiswa, 2-8 ℃, hupenyu hwesherufu mwedzi mitanhatu.
Murairo wekushanda
1. Reaction System (Tora 25μL semuenzaniso)
| Zvikamu | Vhoriyamu (μL) | Final Concentration |
| Lyo-Buffer | 6 | 1* |
| Lyo-Enzyme Mix | 1 | - |
| Lyo-Protectant | 8 | - |
| Primer Mix (10μM) | 1 | 0.1- 1uM |
| Probe Mix (10μM) | 0.5 | 0.05-0.5uM |
| RNA template | 5 | - |
| DEPC H2O | Kusvikira ku25 | - |
2. Optimized Cycling Protocol
1) Standard Cycling Protocol
| Reaction stage | Tembiricha | Nguva | Cycle | |
| 1 | Reverse transcription | 50°Ca | 10min | 1 |
| 2 | Initial denaturation | 95°C | 5min | 1 |
| 3 | Amplification reaction | 95°C | 15sec | 45 kutenderera |
| 60°Cb | 30secc |
2) Fast Cycling Protocol
|
| Reaction stage | Tembiricha | Nguva | Cycle |
| 1 | Reverse transcription | 50°Ca | 2min | 1 |
| 2 | Initial denaturation | 95°C | 2sec | 1 |
| 3 | Amplification reaction | 95°C | 1sec |
45 kutenderera |
| 60°Cb | 13secc |
Cherechedza:
a) Reverse transcription: Tembiricha inogona kusarudza 42°C kana 50°C kwemaminitsi 10-15.
b) Amplification reaction: Iyo tembiricha inogadziriswa zvinoenderana neTm kukosha kweiyo yakagadzirirwa primers.
c)Fluorescence chiratidzo kuwana: Ndapota isa maitiro ekuedza maererano nezvinodiwa zvemudziyo bhuku.
Ruzivo rwehunyanzvi / Ruzivo
| Sravana Sameeralu Serial 4th Hot Start | Yakavakwa-mukati inopisa kutanga |
| Nzira yekuona | Primer-probe yekuona |
| PCR nzira | Imwe nhanho RT-qPCR |
| Rudzi rwemuenzaniso | RNA |
Notes
1. Ichi chigadzirwa ndechekutsvaga kushandiswa chete.
2. Ndokumbira upfeke PPE inodiwa, jasi rakadaro uye magirovhosi, kuve nechokwadi chehutano hwako nekuchengeteka!
