Kayayyaki

Ana amfani da gishirin Tris acetate akai-akai a cikin shirye-shiryen TAE (Tris-acetate-EDTA), wanda aka yi amfani da shi azaman buffer mai gudana kuma a cikin gels agarose.Ana amfani da buffer Tris Acetate-EDTA don DNA agarose gel electrophoresis amma kuma ana amfani dashi don RNA agarose gel electrophoresis mara amfani.DNA mai ɗaure biyu yana ƙoƙarin gudu da sauri a cikin TAE fiye da sauran masu buffer kuma yana iya zama gajiya yayin tsawaita electrophoresis.Buffer wurare dabam dabam ko maye gurbin buffer a lokacin tsawaita electrophoresis na iya magance ƙananan ƙarfin buffer.Ana iya amfani da su a wurare daban-daban don nazarin motsin DNA a cikin bayani.Tun da borate a cikin buffer TBE (Tris-Borate-EDTA Buffer, 10X Powder Pack, sc-296651) shine mai hanawa mai karfi don yawancin enzymes, TAE buffer yana bada shawarar lokacin kallon aikace-aikacen enzymatic don samfurin DNA.Tris acetate gishiri kuma shine buffer tare da babban hankali a cikin gwaje-gwajen ATP tare da luciferase firefly.

Yana amfani da: TAE Gudun buffer shine mafi yawan amfani da buffer don DNA agarose gel electrophoresis, kuma ana amfani dashi don RNA agarose gel electrophoresis na asali.DNA mai ɗaure biyu yana ƙoƙarin motsawa cikin sauri a cikin TAE fiye da sauran masu buffer, amma kuma ya kasa yin iyo saboda raguwar ions buffer a lokacin tsawaita electrophoresis.Keke keken buffer ko musanya buffer yayin tsawaita electrophoresis na iya rama ƙaramin ƙarfin buffer.2 Tsarma madaidaicin buffer TAE don samun buffer 1 mMTAE mai ɗauke da 40 mM Tris acetate da 1 mM EDTA, pH 8.3.Ana iya amfani da buffer 1 mMTAE duka a cikin gels na agarose kuma azaman buffer mai gudana.Don matsakaicin ƙuduri, ana ba da shawarar cewa ƙarfin lantarki da ake amfani da shi ya kasance ƙasa da 5V/cm (nisa tsakanin naúrar naúrar).

Aikace-aikace: TAE Gudun buffer shine mafi yawan amfani da buffer don DNA agarose gel electrophoresis a cikin Gel na Chemicalbook, kuma ana amfani dashi don RNA agarose gel electrophoresis mara amfani.DNA mai ɗaure biyu yana ƙoƙarin motsawa cikin sauri a cikin TAE fiye da sauran masu buffer, amma kuma ya kasa yin iyo saboda raguwar ions buffer a lokacin tsawaita electrophoresis.Keke keken buffer ko musanya buffer yayin tsawaita electrophoresis na iya rama ƙaramin ƙarfin buffer.An narkar da madaidaicin buffer TAE don samun buffer 1 mMTAE mai ɗauke da 40 mM Tris acetate da 1 mM EDTA, pH 8.3.Ana iya amfani da buffer 1 mMTAE duka a cikin gels na agarose kuma azaman buffer mai gudana.Don matsakaicin ƙuduri, ana ba da shawarar cewa ƙarfin lantarki da ake amfani da shi ya kasance ƙasa da 5V/cm (nisa tsakanin naúrar naúrar).

Amfani: A cikin gano ATP tare da firefly luciferase, wannan samfurin shine mafi mahimmancin buffer;gano dauri glutamate.

Ƙunƙarar dauri na [3H]l-glutamic acid zuwa kayan da ba masu karɓa ba.

[3H] L-glutamic acid mai ɗaure ga bututun microfuge da gilashi an bincika su a cikin buffer huɗu.Haɗin bangon baya ga waɗannan kayan ba ya da kyau, amma an ƙara ta ta hanyar centrifugation ko tsotsa a cikin Tris-HCl da Tris-citrate buffer.Wannan ɗaurin ya yi ƙasa da ƙasa ko Lokacin da aka yi amfani da HEPES-KOH, ko Tris-acetate buffer maimakon.[3H] L-glutamate daure zuwa bututun microfuge an hana shi ta L- amma ba D-isomers na glutamate da aspartate ba.DL-2-amino-7-phosphonoheptanoic acid shima bai hana daurin ba.Sauran mahadi waɗanda suka nuna ƙarancin hanawa zuwa matsakaici sune: N-methyl-D-aspartate, qusqualate, L-glutamic acid diethyl ester, N-methyl-L-aspartate, kainate, da 2-amino-4-phosphonobutyrate.An hana dauri ta hanyar barnar kwakwalwar bera.An sami abin daure mai dogaro da furotin [3H] glutamate tare da shirye-shiryen membrane mai daskararre akai-akai lokacin da aka ɗaure a cikin buffer Tris-acetate.Ana ba da shawarar cewa Tris-acetate ko HEPES -KOH buffer yakamata a yi amfani da shi a cikin gwajin glutamate bin ding.Idan an yi amfani da buffer Tris-HCl ko Tris-citrate, yakamata a yi gwajin sarrafawa da ya dace don daidaita bututun microfuge ko matatar fiber gilashi.