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Ikiti ye-PCR yeZilwanyana eziNgqo (ngaphandle kweZixhobo zokuSampula) iProtocol

Inkcazelo yeKit:

Sebenzisa ngokuthe ngqo izicubu zezilwanyana i-lysate njengethempleyithi yokukhulisa i-PCR, ngaphandle kokutsalwa okwahlukileyo kwe-DNA, ngokukhawuleza kunye novakalelo oluphezulu.

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 unonyamezelo oluqinileyo kwi-PCR reaction inhibitors, kwaye unokusebenzisa i-lysate yesampuli ukuba ivavanywe njengethemplate yokukhulisa ngokufanelekileyo kunye nokucacileyo.Le reagent iqulethe Foregene D-Taq DNAPolymerase, dNTPs, MgCl2, i-reaction buffer, i-PCR optimizer kunye ne-stabilizer.

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

Isithinteli AL

I-Foregene Protease

I-6 × i-DNA yokulayisha i-Buffer

Icandelo II

2 × PCR EasyTMXuba

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.

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 EasyTMUmxube ungagcinwa iinyanga ezili-12 kwi -20;ukuba isetyenziswa rhoqo, inokugcinwa ngo-4ukugcinwa kwexesha elifutshane (sebenzisa phezulu kwiintsuku ezili-10).


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