Multiplex Intambwe imwe RT-qPCR Imbere
Ibisobanuro
Injangwe No: HCR5141A
Multiplex Intambwe imwe RT-qPCR Premix ni Multlex igizwe numubare PCR Kit ishingiye kuri RNA nkicyitegererezo.Mubigeragezo, guhinduranya transcript hamwe na PCR yuzuye bikorerwa mumiyoboro imwe, byoroshya imikorere yubushakashatsi no kugabanya ibyago byo kwanduza.Igishushanyo cyihariye cya buffer na enzyme ivanze irashobora gukoreshwa murwego rumwe rwa lyofilize.Igikoresho gikoresha ubushyuhe butarwanya ubushyuhe bwa Transcriptase kugirango ikoreshwe neza ya cDNA ya mbere ikoresheje hotstart Taq DNA Polymerase kugirango yongere umubare.Harimo uburyo bwiza bwo gukora reaction ya buffer, enzymes ivanze nibindi, nibintu bibuza neza kwongera PCR idasanzwe no kongera ubushobozi bwo kongera imbaraga za reaction nyinshi za qPCR byongewemo, bituma florescence nyinshi yongerera imbaraga mugihe harebwa uburyo bwo kongera imbaraga za primers.
Ibigize
| Izina |
| 1. Lyo-Buffer |
| 2. Kuvanga Lyo-Enzyme |
| 3. Kurinda Lyo |
Ubwikorezi bwo gutwara abantuion
Igisubizo: Lyo-buffer kandi ikingira: -25 ~ -15 ℃, ubuzima bwo kubaho ni umwaka 1.
B: Lyo-enzyme ivanze, 2-8 ℃, ubuzima bwo kubaho ni amezi 6.
Amabwiriza yo gukora
1. Sisitemu yo Kwitwara (Fata 25μL nkurugero)
| Ibigize | Umubumbe (μL) | Kwibanda kwanyuma |
| Lyo-Buffer | 6 | 1* |
| Lyo-Enzyme ivanze | 1 | - |
| Lyo-Kurinda | 8 | - |
| Kuvanga Primer (10μM) | 1 | 0.1- 1uM |
| Kuvanga Ubushakashatsi (10μM) | 0.5 | 0.05-0.5uM |
| Inyandikorugero ya RNA | 5 | - |
| DEPC H2O | Kugera kuri 25 | - |
2. Porotokole nziza yo gusiganwa ku magare
1) Porotokole isanzwe yo gusiganwa ku magare
| Icyiciro cya reaction | Ubushyuhe | Igihe | Ukuzenguruka | |
| 1 | Guhindura inyandiko | 50 ° C.a | 10min | 1 |
| 2 | Gutandukana kwambere | 95 ° C. | 5min | 1 |
| 3 | Amplification reaction | 95 ° C. | 15sec | Inzinguzingo 45 |
| 60 ° C.b | 30secc |
2) Porotokole Yihuta Yamagare
|
| Icyiciro cya reaction | Ubushyuhe | Igihe | Ukuzenguruka |
| 1 | Guhindura inyandiko | 50 ° C.a | 2min | 1 |
| 2 | Gutandukana kwambere | 95 ° C. | 2sec | 1 |
| 3 | Amplification reaction | 95 ° C. | 1sec |
Inzinguzingo 45 |
| 60 ° C.b | 13secc |
Icyitonderwa:
a) Subiza inyuma transcript: Ubushyuhe burashobora guhitamo 42 ° C cyangwa 50 ° C muminota 10-15.
b) Amplification reaction: Ubushyuhe bwahinduwe ukurikije agaciro ka Tm ya primers yagenewe.
c)Fluorescence ikimenyetso kugura: Nyamuneka shiraho uburyo bwo kugerageza ukurikije ibisabwa byaimfashanyigisho.
Amakuru ya tekiniki / Ibisobanuro
| Intangiriro ishyushye | Byubatswe-bishyushye gutangira |
| Uburyo bwo kumenya | Kugaragaza Primer-probe |
| Uburyo bwa PCR | Intambwe imwe RT-qPCR |
| Ubwoko bw'icyitegererezo | RNA |
Inyandiko
1. Iki gicuruzwa nikoreshwa mubushakashatsi gusa.
2. Nyamuneka wambare PPE ikenewe, ikote rya laboratoire hamwe na gants, kugirango ubuzima bwawe n'umutekano！
