I-Zebra Fish Direct PCR Kit Direct PCR Lysis Reagent(zebrafish)
Iinkcazelo
Le mveliso isebenzisa inkqubo eyodwa ye-lysis buffer ukukhulula ngokukhawuleza i-DNA ye-genomic kwi-zebrafish kunye nezinye izihlunu zentlanzi zamanzi ahlambulukileyo, ama-fins omsila okanye iisampuli zamaqanda eentlanzi kwiimpendulo ze-PCR, ngoko ke ifaneleka ngakumbi kuvavanyo lofuzo olukhulu.Inkqubo yokukhulula i-DNA ye-genomic kwi-lysis buffer igqityiwe ngaphakathi kwe-10-30 imizuzu kwi-65 ° C.Azikho ezinye iinkqubo ezifana neprotheyini kunye nokususwa kwe-RNA ezifunekayo, kwaye umkhondo okhutshiweyo we-DNA ungasetyenziswa njenge template ye-PCR reaction.
2×PCR EasyTMUmxube unonyamezelo oluqinileyo kwi-PCR reaction inhibitors, kwaye unokusebenzisa i-lysate yesampuli ukuba ivavanywe njengethemplate yokukhulisa ngokufanelekileyo kunye nokucacileyo.Le reagent iqulethe ForegeneD-Taq DNAPolymerase, dNTPs, MgCl2, i-reaction buffer, i-PCR optimizer kunye ne-stabilizer.Isetyenziswe ngokubambisana ne-lysis buffer inokukhawuleza kwaye ibone iisampulu ngokulula, kwaye ineempawu zobuntununtunu obuphezulu, ukuchaneka okunamandla, kunye nokuzinza okulungileyo.
I-D-Taq DNA polymerase yipolymerase ye-DNA ephuhliswe ngokukodwa yi-Foregene ukwenzela ukuphendula ngokuthe ngqo kwe-PCR.I-D-Taq DNA polymerase inonyamezelo oluqinileyo kwiintlobo ezahlukeneyo ze-PCR reaction inhibitors, kwaye inokwandisa ngokufanelekileyo izixa ze-DNA kwiinkqubo ezahlukeneyo zokusabela ezintsonkothileyo, kunye nesantya sokukhulisa sinokufikelela kwi-2Kb / min, efanelekileyo kakhulu kwi-Direct PCR reaction.
Iinkcukacha
50×20µl rxns, 200×20µl rxns, 500×20µl rxns, 2000×20µl rxns
Amacandelo ekhithi
Icandelo I | Buffer FP |
I-Foregene Protease | |
I-6 × i-DNA yokulayisha i-Buffer | |
Icandelo II | 2×PCR EasyTMXuba |
Imiyalelo |
Iimpawu&izinto eziluncedo
■Akukho mfuneko yokucocwa kwe-DNA okuchitha ixesha kunye nexabiso eliphezulu.
∎ Imfuno yesampulu incinci, kancinane njenge-1mg ye-caudal fin yentlanzi okanye amaqanda entlanzi ali-10 anokuhlolwa.
∎ Akukho nyango lukhethekileyo olunjengokusila nokutyumza olufunekayo, kwaye utyando lulula.
■ Inkqubo ye-PCR ephuculweyo yenza ukuba i-PCR ibe nezinto ezicacileyo eziphezulu kunye nokunyamezelana okunamandla kwii-PCR reaction inhibitors.
Iiparamitha zeKit
Isicelo: Iintlanzi zeZebra kunye nezinye iintlanzi zamanzi ahlambulukileyo.
I-DNA ekhutshwe kwisampuli ye-lysis: isetyenziswe kuphela njenge-template ye-PCR.
Ikiti ingasetyenziselwa ezi njongo zilandelayo: ukuchongwa kwe-transgenosis, i-genotyping yezilwanyana, njl.
Ukuhamba komsebenzi
Ugcino kunye nobomi beShelf
Inxalenye I yale khithi kufuneka igcinwe kwi-2-8℃.
I-reagent Buffer FP inokugcinwa iinyanga ezili-12 phantsi kweemeko ezomileyo;inokugcinwa kwi-2-8℃ ukuze igcinwe ixesha elide.
v IReagent Foregene Protease inefomula eyodwa.Ukuqinisekisa umsebenzi wayo kunye nokuzinza, nceda uyigcine kwi-4 ℃ iinyanga ezili-12.
vReagent 6×DNA Loading Buffer ingagcinwa kwi 4℃ okanye -20℃ ixesha elide.
Icandelo II lale khithi kufuneka ligcinwe kwi -20 ℃.
I-vReagent 2 × PCR Easy TM Mix, ukuba isetyenziswe rhoqo, ingaphinda igcinwe kwi-4 ℃ yokugcina ixesha elifutshane (sebenzisa kwiintsuku ze-10).