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I-Animal Tissue Direct PCR kit-UNG (ngaphandle kweSampuli yeZixhobo) iProtocol

Inkcazelo yeKit:

Sebenzisa ngokuthe ngqo izicubu zezilwanyana i-lysate njengethempleyithi yokukhulisa i-PCR, ngaphandle kokutsalwa okwahlukileyo kwe-DNA, ngokukhawuleza kunye novakalelo oluphezulu.Kongezwe inkqubo ye-UNG yokuchasana nokungcoliseka kongcoliseko ukuze kupheliswe iimpawu zobuxoki.

Akukho mfuneko yokucocwa kwe-DNA okuchitha ixesha kunye nexabiso eliphezulu.

Imfuno yesampulu incinci, thatha nje imbewu enye.

Akukho nyango lukhethekileyo olunjengokusila kunye nokutyumza olufunekayo, kwaye umsebenzi ulula.

Inkqubo ye-PCR elungiselelweyo yenza ukuba i-PCR ibe nokucaciswa okuphezulu kunye nokunyamezelana okunamandla kwi-PCR reaction inhibitors.

amandla angaphambili


Iinkcukacha zeMveliso

Iithegi zeMveliso

Iinkcazelo

Le khithi isebenzisa inkqubo ye-lysis buffer ekhethekileyo ukukhulula ngokukhawuleza i-DNA ye-genomic kwiisampuli zezicubu zezilwanyana ukwenzela ukusabela kwe-PCR, ngoko ke ifaneleke ngokukodwa kuvavanyo olukhulu lwemfuzo.

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 (UNG) usebenzisa i-dUTP endaweni ye-dTTP kwisiseko se-2×PCR EasyTMHlanganisa, kwaye ungeze i-enzyme ye-UNG (i-Uracil-N-glycosylase) enokunciphisa i-template equkethe i-dUTP ngexesha elifanayo.Ngaphambi kokuphendula kwe-PCR, i-enzyme ye-UNG isetyenziselwa ukuthotywa kwemveliso ye-PCR equkethe i-uracil.I-enzyme ye-UNG ayiyi kuba nayiphi na impembelelo kwi-template engenawo i-uracil, ngaloo ndlela iqinisekisa ukuchaneka kunye nokuchaneka kwe-amplification kunye nokuthintela ukuba kungenzeka ukuba ukungcola kweemveliso ze-PCR ngexesha lovavanyo olukhulu lofuzo.

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.

Amacandelo ekhithi

Icandelo I

Isithinteli AL

I-Foregene Protease

I-6 × i-DNA yokulayisha i-Buffer

Icandelo II

2×PCR EasyTMUmxube(UNG)

Iimpawu&izinto eziluncedo

Akukho xesha lidla kwaye libiza ukucocwa kweDNA

∎ Imfuno yesampulu incinci, kancinane njenge-5mg inyama yesilwanyana inokuvavanywa.

∎ 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.

■ Inkqubo yePCR yokuchasana nongcoliseko 2×PCR EasyTMUkuxuba (UNG) kuphelisa ngokufanelekileyo ukungcola okubangelwa yimveliso ye-PCR kwaye iqinisekisa ukuchaneka kunye nokuchaneka kokukhulisa.

Iiparamitha zeKit

Isicelo: iintlobo ezahlukeneyo zezicubu zezilwanyana.

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

wk-flw-isilwanyana kwi-PCR ngqo

Ugcino kunye nobomi beShelf

Inxalenye I yale khithi kufuneka igcinwe kwi-2-8.

I-reagent Buffer AL inokugcinwa iinyanga ezili-12 phantsi kweemeko ezomileyo;inokugcinwa kwi-2-8yokugcina ixesha elide.

 I-Reagent Foregene Protease inefomula eyodwa.Ukuqinisekisa umsebenzi wayo kunye nokuzinza, nceda uyigcine ku-4iinyanga ezili-12.

 I-Reagent 6×I-DNA Loading Buffer inokugcinwa kwi-4okanye -20ixesha elide.

Inxalenye II yale khithi kufuneka igcinwe kwi -20.

 I-Reagent 2×PCR EasyTMMix(UNG), ukuba isetyenziswa rhoqo, inokugcinwa ngo4ukugcinwa kwexesha elifutshane (sebenzisa phezulu kwiintsuku ezili-10).


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