Ikhithi Yokuhlukanisa I-DNA Yamanzi I-DNA Isizinda kanye Nekhithi Yokuhlanza Yamanzi
Incazelo yekhithi:
Hlanza ngokushesha i-DNA ye-genomic yekhwalithi ephezulu kuma-microorganisms kumasampula amanzi ahlukahlukene.
◮Akukho ukungcoliswa kwe-RNase:Ikholomu Ye-DNA Kuphela ehlinzekwa ikhithi yenza kube nokwenzeka ukususa i-RNA ku-DNA ye-genomic ngaphandle kokwengeza i-RNase phakathi nokuhlolwa, ukuvimbela ilabhorethri ukuthi ingangcoliswa i-RNase yangaphandle.
◮Isivinini esisheshayo:I-Foregene Protease inomsebenzi ophezulu kunama-protease afanayo, futhi igaya amasampula ezicubu ngokushesha;ukusebenza kulula, futhi ukusebenza kwe-genomic DNA extraction kungaqedwa phakathi kwemizuzu engu-20-80.
◮Kulula:I-centrifugation yenziwa ekamelweni lokushisa, futhi asikho isidingo se-centrifugation yezinga lokushisa eliphansi elingu-4°C noma imvula ye-ethanol ye-DNA.
◮Ukuphepha:Akukho kukhishwa kwe-organic reagent edingekayo.
◮Ikhwalithi ephezulu:I-DNA ye-genomic ekhishiwe inezicucu ezinkulu, ayikho i-RNA, ayikho i-RNase, nokuqukethwe kwe-ion ephansi kakhulu, okungahlangabezana nezidingo zokuhlola okuhlukahlukene.
◮Isistimu ye-Micro-elution:Ingakhuphula ukugxila kwe-genomic DNA, elungele ukutholwa komfula noma ukuhlolwa.
Imininingwane
50 Preps
-Lysozyme: enzymatic hydrolyze udonga lwamaseli amagciwane aqondile.
-I-Buffer TE: isetshenziselwa ukulungisa isisombululo se-100mg/ml Lysozyme futhi inikeze imvelo ye-lysozyme enzymatic hydrolysis.
-Buffer SG1 & Buffer SG2: Nikeza isampula yendawo yokugaya ye-protease.
-I-Forgene Protease: Amasampula e-Enzymatic hydrolyze endaweni ye-protease enzymatic ukuze akhulule i-genomic DNA.
-Buffer SG3: Yenza i-Foregene Protease isebenze futhi unikeze indawo yokulayisha i-DNA.
-Buffer SG4: Isengezo sokuhlinzeka ngemvelo yokulayisha i-DNA.
-Buffer PW: Susa ukungcola okufana namaprotheni ne-RNA ku-DNA.
I-Buffer WB: Susa ama-ion kasawoti asele ku-DNA.
-Buffer EB: susa i-DNA kulwelwesi lwekholomu yokuhlanza.
-Ikholomu ye-DNA kuphela: khangisa ngokukhethekile i-DNA ye-genomic ku-lysate.
Izingxenye zekhithi
| Isilondolozi SG1 |
| Isilondolozi SG2 |
| Isilondolozi SG3 |
| Isilondolozi SG4 |
| Isilondolozi se-PW |
| Isilondolozi se-WB |
| Isilondolozi EB |
| Isilondolozi TE |
| I-Foregene Protease |
| I-Lysozyme |
| Ikholomu Ye-DNA Kuphela |
| Iziyalezo |
Izici&izinzuzo
-Akukho ukungcola kwe-RNase: Ikholomu Ye-DNA Kuphela ehlinzekwa ikhithi yenza kube nokwenzeka ukususa i-RNA ku-DNA ye-genomic ngaphandle kwe-RNase eyengeziwe phakathi nokuhlolwa, ukugwema ilabhorethri ukuthi ingangcoliswa i-RNase yangaphandle.
-Isivinini esisheshayo: Ukusebenza kulula, futhi ukusebenza kwe-genomic DNA extraction yamanzi kungaqedwa kungakapheli imizuzu engama-40.
-Kulula: I-centrifugation yenziwa ekamelweni lokushisa, futhi asikho isidingo se-centrifugation yezinga lokushisa eliphansi elingu-4°C noma imvula ye-ethanol ye-DNA.
-Ukuphepha: asikho isidingo sokukhishwa kwe-organic reagent.
-Ikhwalithi ephezulu: Izingcezu ze-genomic DNA ezikhishiwe zinkulu, azikho i-RNA, azinayo i-RNase, nokuqukethwe kwe-ion okuphansi kakhulu, okungahlangabezana nezidingo zokuhlolwa okuhlukahlukene.
Isicelo sekhithi
Ilungele ukuhlanzwa kwe-DNA ye-genomic yamasampula alandelayo: amanzi echibi le-aquaculture, amanzi echibi, amanzi ampompi, amanzi echibi, amanzi omfula, amanzi echibi le-lotus namanye amasampula.
Ukuhamba komsebenzi
Isitoreji kanye Nempilo yeshelufu
-Le kit ingagcinwa izinyanga ezingu-12 ngaphansi kwezimo ezomile ekamelweni lokushisa (15-25 ° C), uma idinga ukugcinwa isikhathi eside, ingagcinwa ku-2-8 ° C.
Qaphela: Uma igcinwe ezingeni lokushisa eliphansi, isixazululo sithambekele ekubeni nemvula.Ngaphambi kokusetshenziswa, qiniseka ukuthi ubeke ikhambi kukhithi endaweni yokushisa yasekamelweni isikhathi esithile.Uma kunesidingo, yishise ngaphambili ngo-37°C okugeza ngamanzi imizuzu eyi-10 ukuncibilikisa imvula, futhi uyixube ngaphambi kokusetshenziswa.
-Isixazululo se-Forgene Protease sinefomula eyingqayizivele, esebenzayo uma igcinwe ekamelweni lokushisa isikhathi eside (izinyanga ezingu-3);uma igcinwe ku-4 ℃, umsebenzi wayo nokuzinza kuzoba ngcono, ngakho-ke kunconywa ukuyigcina ku-4 ℃, khumbula ukuthi ungayigcini ku -20 ℃.
-I-powder eyomile i-Lysozyme igcinwa ku -20 ° C;isisombululo se-Lysozyme esilungisiwe sihlukaniswe izingxenye ezincane futhi sigcinwe ku -20 ° C.
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Ikhithi yokuhlukanisa i-DNA yamahhala
