Tsiaj Tissue Direct PCR cov khoom siv-UNG (tsis suav cov cuab yeej siv) raws tu qauv
Cov lus piav qhia
Cov khoom siv no siv cov txheej txheem lysis tsis zoo kom sai sai tso cov genomic DNA los ntawm cov tsiaj cov ntaub so ntswg kuaj rau PCR cov tshuaj tiv thaiv, yog li nws yog tshwj xeeb tshaj yog haum rau loj-scale kuaj caj ces.
Cov txheej txheem ntawm kev tso genomic DNA los ntawm lysis tsis yog ua tiav hauv 10-30 feeb ntawm 65 ° C.Tsis muaj lwm yam txheej txheem xws li protein thiab RNA tshem tawm, thiab cov kab DNA tso tawm tuaj yeem siv los ua qauv rau PCR cov tshuaj tiv thaiv.
2 × PCR Ib qho yooj yimTMMix (UNG) siv dUTP hloov dTTP raws li 2 × PCR Yooj YimTMSib tov, thiab ntxiv UNG enzyme (Uracil-N-glycosylase) uas tuaj yeem degrade cov qauv uas muaj dUTP tib lub sijhawm.Ua ntej cov tshuaj tiv thaiv PCR, UNG enzyme yog siv los degrade cov khoom PCR uas muaj uracil.Lub UNG enzyme yuav tsis muaj kev cuam tshuam rau tus qauv uas tsis muaj uracil, yog li ua kom muaj qhov tshwj xeeb thiab qhov tseeb ntawm kev nthuav dav thiab tiv thaiv qhov ua tau ntawm PCR cov khoom sib kis thaum lub sij hawm loj-scale kuaj caj ces.
D-Taq DNA polymerase yog DNA polymerase tshwj xeeb uas tsim los ntawm Foregene rau cov tshuaj PCR ncaj qha.D-Taq DNA polymerase muaj kev ua siab ntev rau ntau yam PCR cov tshuaj tiv thaiv, thiab tuaj yeem ua kom muaj txiaj ntsig zoo ntawm DNA hauv ntau cov tshuaj tiv thaiv nyuaj, thiab qhov kev nthuav dav tuaj yeem ncav cuag 2Kb / min, uas yog tshwj xeeb tshaj yog haum rau Direct PCR cov tshuaj tiv thaiv.
Cov khoom siv
Ntu I | Buffer AL |
Foregene Protease | |
6 × DNA Loading Buffer | |
Ntu II | 2 × PCR Ib qho yooj yimTMMix (UA) |
Nta & qhov zoo
■Tsis siv sijhawm ntev thiab kim DNA purification
■ Cov qauv xav tau me me, tsawg li 5mg cov ntaub so ntswg tsiaj tuaj yeem kuaj tau.
■ Tsis muaj kev kho tshwj xeeb xws li kev sib tsoo thiab kev sib tsoo, thiab kev ua haujlwm yooj yim.
■ Optimized PCR system ua rau PCR muaj qhov tshwj xeeb dua thiab muaj zog ua siab ntev rau PCR cov tshuaj tiv thaiv cov tshuaj tiv thaiv.
■ Anti-pollution PCR system 2×PCR Ib qho yooj yimTMMix (UNG) zoo tshem tawm cov pa phem los ntawm PCR cov khoom thiab xyuas kom meej qhov tseeb thiab qhov tseeb ntawm amplification.
Cov khoom siv parameter
Daim ntawv thov: ntau yam tsiaj cov ntaub so ntswg.
DNA tso tawm los ntawm cov qauv lysis: tsuas yog siv los ua PCR template.
Cov khoom siv tuaj yeem siv rau lub hom phiaj hauv qab no: kev txheeb xyuas ntawm transgenosis, tsiaj genotyping, thiab lwm yam.
Kev khiav haujlwm
Cia thiab Txee lub neej
Ntu I ntawm cov khoom siv no yuav tsum muab khaws cia ntawm 2-8℃.
Lub reagent Buffer AL tuaj yeem khaws cia rau 12 lub hlis nyob rau hauv qhuav qhuav;nws tuaj yeem khaws cia ntawm 2-8℃rau ntev cia.
Reagent Foregene Protease muaj cov qauv tshwj xeeb.Txhawm rau kom ntseeg tau tias nws cov haujlwm thiab kev ruaj ntseg, thov khaws cia ntawm 4℃rau 12 lub hlis.
Reagent 6×DNA Loading Buffer tuaj yeem khaws cia ntawm 4℃los yog -20℃ntev.
Ntu II ntawm cov khoom siv no yuav tsum muab khaws cia ntawm -20℃.
Reagent 2×PCR Ib qho yooj yimTMMix (UNG), yog tias nws siv ntau zaus, nws tuaj yeem khaws cia ntawm 4℃rau lub sij hawm luv luv (siv li ntawm 10 hnub).