Ibicuruzwa

Umunyu wa Tris acetate ukoreshwa kenshi mugutegura buffer ya TAE (Tris-acetate-EDTA), ikoreshwa nka buffer ikora no muri geles ya agarose.Tris Acetate-EDTA buffer ikoreshwa kuri ADN agarose gel electrophorei ariko ikoreshwa no muburyo budasanzwe bwa RNA agarose gel electrophorei.ADN ikubye kabiri ikunda gukora byihuse muri TAE kuruta iyindi miyoboro kandi irashobora kunanirwa mugihe cya electrophorei yagutse.Gukwirakwiza kwa buffer cyangwa gusimbuza buffer mugihe kinini cya electrophoreis irashobora gukemura ubushobozi bwo hasi.Birashobora gukoreshwa mubitekerezo bitandukanye kugirango bige kugenda kwa ADN mugukemura.Kubera ko borate muri buffer ya TBE (Tris-Borate-EDTA Buffer, 10X Powder Pack, sc-296651) ni inhibitor ikomeye kuri enzymes nyinshi, buffer ya TAE irasabwa mugihe ureba progaramu ya enzymatique ya sample ya ADN.Umunyu wa Tris acetate nawo ni buffer ifite sensibilité nyinshi muri ATP isuzuma hamwe na luciferase yumuriro.

Imikoreshereze: TAE ikoresha buffer niyo ikoreshwa cyane muri ADN agarose gel electrophorei, kandi ikoreshwa no muri RNA agarose gel electrophorei.ADN ikubye kabiri ikunda kugenda byihuse muri TAE kuruta iyindi bufferi, ariko kandi ikananirwa koga bitewe no kugabanuka kwa ion ion mugihe cya electrophoreze igihe kirekire.Umukino wo gusiganwa ku magare cyangwa guhinduranya mugihe cya electrophoreis igihe kirekire birashobora kwishyura ubushobozi buke bwo hasi.2 Koresha buffer ya TAE yibanze kugirango ubone buffer 1 mMTAE irimo 40 mM Tris acetate na 1 mM EDTA, pH 8.3.Buffer ya mMTAE 1 irashobora gukoreshwa haba muri geles ya agarose no nka buffer ikora.Kugirango bikemuke ntarengwa, birasabwa ko voltage ikoreshwa iba munsi ya 5V / cm (intera iri hagati ya electrode).

Gushyira mu bikorwa: TAE ikoresha buffer nizo zikoreshwa cyane muri ADN agarose gel electrophorei muri gel ya Chemicalbook, kandi ikoreshwa no mu kudatandukanya RNA agarose gel electrophorei.ADN ikubye kabiri ikunda kugenda byihuse muri TAE kuruta iyindi bufferi, ariko kandi ikananirwa koga bitewe no kugabanuka kwa ion ion mugihe cya electrophoreze igihe kirekire.Umukino wo gusiganwa ku magare cyangwa guhinduranya mugihe cya electrophoreis igihe kirekire birashobora kwishyura ubushobozi buke bwo hasi.Buffer yibanze ya TAE yahinduwe kugirango ibone buffer 1 mMTAE irimo 40 mM Tris acetate na mM 1 EDTA, pH 8.3.Buffer ya mMTAE 1 irashobora gukoreshwa haba muri geles ya agarose no nka buffer ikora.Kugirango bikemuke ntarengwa, birasabwa ko voltage ikoreshwa iba munsi ya 5V / cm (intera iri hagati ya electrode).

Gukoresha: Mugushakisha ATP hamwe na luciferase yumuriro, iki gicuruzwa nicyo kintu cyoroshye cyane;glutamate binding detection.

Guhinduranya kwimura [3H] l-glutamic aside kubikoresho bitakira.

[3H] L-glutamic aside ihuza imiyoboro ya microfuge nikirahure byakorewe ubushakashatsi muri bffers enye.Amavu n'amavuko guhuza ibyo bikoresho ntibyari byemewe, ariko yongerewe na centrifugation cyangwa guswera muri Tris-HCl na Tris-citrate buffer.Uku guhambira kwari gake cyane cyangwa Iyo HEPES-KOH, cyangwa buffer ya Tris-acetate yakoreshejwe aho.[3H] L-glutamate ihuza imiyoboro ya microfuge yabujijwe na L- ariko ntabwo D-isomers ya glutamate na aspartate.DL-2-amino-7-fosifoneheptanoic aside nayo ntiyabujije guhambira.Ibindi bikoresho byerekanaga ko bibujijwe kugera ku rugero ruto: N-methyl-D-aspartate, quisqualate, L-glutamic aside diethyl ester, N-methyl-L-aspartate, kainate, na 2-amino-4 -phosphonobutyrate.Guhambira byahagaritswe n'imbeba zo mu bwonko.Guhuza poroteyine [3H] glutamate yabonetse hamwe nogutegura inshuro nyinshi gukonjesha mugihe cyo guhambira byakorewe muri buffer ya Tris-acetate.Birasabwa ko Tris-acetate cyangwa HEPES -KOH buffer igomba gukoreshwa muri glutamate bin ding assay.Niba hakoreshejwe Tris-HCl cyangwa Tris-citrate buffer, hagomba gukorwa igerageza rikwiye ryo gukosora kugirango uhuze imiyoboro ya microfuge cyangwa ibirahuri bya fibre.