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isibhengezo_sekhasi

I-Zebra Fish Direct PCR Kit Direct PCR Lysis Reagent(i-zebrafish)

Incazelo yekhithi:

Sebenzisa ngokuqondile i-zebrafish lysate njengesifanekiso sokukhulisa i-PCR, ngaphandle kokukhipha okuhlukile kwe-DNA, ukuzwela okusheshayo nokuphezulu.

Akukho ukuhlanzwa kwe-DNA okuthatha isikhathi nokubizayo okudingekayo.

Isidingo sesampula sincane, thatha nje imbewu eyodwa.

Azikho izindlela zokwelapha ezikhethekile ezinjengokugaya nokuchoboza okudingekayo, futhi ukuhlinzwa kulula.

Isistimu ye-PCR ethuthukisiwe yenza i-PCR ibe nokucaciswa okuphezulu nokubekezelela okunamandla kuma-PCR reaction inhibitors.

amandla angaphambili


Imininingwane Yomkhiqizo

Omaka bomkhiqizo

Izincazelo

Lo mkhiqizo usebenzisa isistimu ye-lysis buffer eyingqayizivele ukuze ukhulule ngokushesha i-DNA ye-genomic ku-zebrafish nezinye izicubu zezinhlanzi zasemanzini ahlanzekile, amaphiko omsila noma amasampula amaqanda ezinhlanzi zokusabela kwe-PCR, ngakho-ke ifaneleka ngokukhethekile ukuhlolwa kofuzo okukhulu.Inqubo yokukhipha i-DNA ye-genomic ku-lysis buffer iqedwa phakathi kwemizuzu eyi-10-30 ku-65°C.Azikho ezinye izinqubo ezidingekayo njengokususwa kwamaprotheni kanye ne-RNA, futhi i-trace DNA ekhishiwe ingasetshenziswa njengesifanekiso sokusabela kwe-PCR.

2×PCR KululaTMI-Mix inokubekezelela okuqinile kwe-PCR reaction inhibitors, futhi ingasebenzisa i-lysate yesampula ukuze ihlolwe njengesifanekiso sokukhulisa kahle nokuqondile.Lesi sikhungo siqukethe i-ForegeneD-Taq DNAPolymerase, i-dNTPs, i-MgCl2, isigcinalwazi sokusabela, i-PCR optimizer nesiqinisi.Isetshenziswa ngokuhambisana ne-lysis buffer ingakwazi ukubona ngokushesha futhi kalula amasampula, futhi inezici zokuzwela okuphezulu, ukucaciswa okuqinile, nokuzinza okuhle.

I-D-Taq DNA polymerase iyi-polymerase ye-DNA ethuthukiswe ngokukhethekile yi-Foregene ukuze kutholakale ukusabela okuqondile kwe-PCR.I-D-Taq DNA polymerase inokubekezelela okunamandla ezinhlobonhlobo ze-PCR reaction inhibitors, futhi ingakhulisa kahle amanani omkhondo we-DNA ezinhlelweni ezihlukene zokusabela eziyinkimbinkimbi, futhi isivinini sokukhulisa singafinyelela ku-2Kb/min, okulungele ikakhulukazi ukusabela kwe-Direct PCR.

Imininingwane

50×20µl rxns, 200×20µl rxns, 500×20µl rxns, 2000×20µl rxns

Izingxenye zekhithi

Ingxenye I

I-Buffer FP

I-Foregene Protease

I-6× I-DNA Yokulayisha Ibhafa

Ingxenye II

2×PCR KululaTMHlanganisa

Iziyalezo

Izici&izinzuzo

Akukho ukuhlanzwa kwe-DNA okuthatha isikhathi nokubizayo okudingekayo.

■ Isidingo sesampula sincane, njengoba nje kungahlolwa i-fish caudal fin engu-1mg noma amaqanda ayi-10 ezinhlanzi.

■ Azikho izindlela zokwelapha ezikhethekile ezinjengokugaya nokuchoboza okudingekayo, futhi ukuhlinzwa kulula.

■ Uhlelo lwe-PCR olulungiselelwe lenza i-PCR ibe nokucaciswa okuphezulu nokubekezelela okuqinile kuma-PCR reaction inhibitors.

Imingcele yekhithi

Isicelo: Izinhlanzi ze-Zebra nezinye izinhlanzi zamanzi ahlanzekile.

I-DNA ekhishwe kusampula ye-lysis: isetshenziswa kuphela njengesifanekiso se-PCR.

Ikhithi ingasetshenziselwa lezi zinhloso ezilandelayo: ukuhlonza i-transgenosis, i-genotyping yezilwane, njll.

Ukugeleza komsebenzi

wk-flw-zebra-fish-direct-PCR

Isitoreji kanye Nempilo yeshelufu

Ingxenye I yale kit kufanele igcinwe ku-2-8℃.

I-reagent Buffer FP ingagcinwa izinyanga eziyi-12 ngaphansi kwezimo ezomile;ingagcinwa ku-2-8℃ ukuze igcinwe isikhathi eside.

v I-Reagent Forgene Protease inefomula eyingqayizivele.Ukuqinisekisa ukusebenza kwayo nokuzinza, sicela uyigcine ku-4℃ izinyanga eziyi-12.

I-vReagent 6×DNA Loading Buffer ingagcinwa ku-4℃ noma -20℃ isikhathi eside.

Ingxenye II yale khithi kufanele igcinwe ku -20 ℃.

I-vReagent 2×PCR EasyTM Mix, uma isetshenziswa njalo, ingabuye igcinwe ku-4℃ ukuze igcinwe isikhathi esifushane (isetshenziswe phakathi kwezinsuku eziyi-10).


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