Multiplex Intambwe imwe RT-qPCR Premix-UNG
Injangwe No: HCB5145A
Multiplex Intambwe imwe RT-qPCR Probe Kit (UDG Plus) nigikoresho cyinshi cya PCR kitwa RNA gishingiye kuri RNA nkicyitegererezo.Mugihe cyibigeragezo, transcription transcription hamwe na PCR yuzuye byakorewe mumiyoboro imwe, byoroshya imikorere yubushakashatsi kandi bigabanya ibyago byo kwanduza.Muri iki gikoresho, umurongo wa mbere cDNA washyizwemo neza na Transversase yihanganira ubushyuhe kandi wongerewe ubwinshi na HotStart Tag ADN Polymerase.Igikoresho kirimo cyane cyane MP ya buffer nziza, enzymes ivanze, nibindi. Igisubizo cya buffer kimaze kubamo Mg2+na DNTP.Byongeye kandi, ibintu bishobora kubuza neza kwongera kwa PCR kutari kwihariye no kunoza imikorere ya reaction nyinshi za qPCR byongeweho, bishobora kwemeza imikorere ya amplification kandi bigakorwa kugeza kuri reaction nyinshi.Sisitemu ya DUTP / UDG yongeweho kugirango irinde neza ibyago byo kwanduza aerosol.
Ibigize
1. Buffer
2. Kuvanga Enzyme
Ibisobanuro
| Intangiriro ishyushye | Byubatswe-bishyushye gutangira |
| Uburyo bwo kumenya | Kugaragaza Primer-probe |
| Uburyo bwa PCR | Intambwe imwe RT-qPCR |
| Polymerase | Taq polymerase |
| Ubwoko bw'icyitegererezo | ADN |
Ububiko
Ibicuruzwa byoherejwe hamwe na barafu yumye kandi birashobora kubikwa kuri -25 ~ -15 ℃ kumwaka 1.Igomba kwirinda kenshigukonjesha.Birasabwa kuzigama ukundi.
Amabwiriza
1.Igisubizo Sisitemu
| Ibigize | Umubumbe (μL) | Kwibanda kwanyuma |
| 2 × Depite Buffer | 12.5 | 1 × |
| Enzyme Kuvanga | 1 | - |
| Primer / Probe ivanze (2.5 μM) | 3 | 0.3μM |
| Inyandikorugero RNA | 1-10 | - |
| RNase Ubuntu H2O | kugeza 25 | - |
Inyandiko:
Witondere kuvanga neza mbere yo gukoresha, irinde ibibyimba byinshi biterwa no kunyeganyega gukabije.
a.Kwibanda kwa primer: Kwivanga kwa primer harimo na multiplex primer, ukurikije uko ibintu bimeze neza primer yibanze wenda hagati ya 0.l na 1.0μM.
b.Kwibanda kuri probe: Kuvanga probe harimo na multiplex probe iranga itandukaniro rya fluorescent group, ukurikije uko ibintu byifashe neza probe wenda hagati ya 0.05 na 0.5μM.
c.Kwiyoroshya kwicyitegererezo: qPCR irakomeye cyane kandi birasabwa kugabanya inyandikorugero.Igenzura Ct agaciro karakwiriye hagati ya 20 na 35.
d.Gutegura sisitemu: Nyamuneka tegura mumeza ya ultra isukuye ikora, hamwe na pipettor na reaction ya reaction idafite ibisigazwa bya nuclease;birasabwa gukoresha umutwe wimbunda hamwe nibintu byungurura.Irinde kwanduza umusaraba no kwanduza aerosol.
2. Amagare mezaPorotokole
| Ukuzenguruka intambwe | Ubushuhe. | Igihe | Amagare |
| Guhindura inyandiko | 50 ℃a | 20min | 1 |
| Intangiriro-gutandukana | 95 ℃ | 5min | 1 |
| Amplification reaction | 95 ℃ | 15sec |
40-45 |
| 60 ℃ b | 30sec c |
Inyandiko:
a.Guhindura inyandiko: Ubushyuhe burashobora guhitamo 42 ° C cyangwa 50 ° C.
b.Amplification reaction: Ubushyuhe burahindurwa ukurikije agaciro ka Tm ya primers yagenewe.
c.Kugura ibimenyetso bya Fluorescence: Nyamuneka shiraho uburyo bwo kugerageza ukurikije ibisabwa nigitabo gikubiyemo ibikoresho.
Inyandiko
Nyamuneka wambare PPE ikenewe, nka kote ya laboratoire na gants, kugirango umenye ubuzima bwawe n'umutekano.
