Zogulitsa

Mchere wa Tris acetate umagwiritsidwa ntchito mobwerezabwereza pokonza buffer ya TAE (Tris-acetate-EDTA), yomwe imagwiritsidwa ntchito ngati buffer yothamanga komanso mu agarose gels.Tris Acetate-EDTA buffer imagwiritsidwa ntchito pa DNA agarose gel electrophoresis koma imagwiritsidwanso ntchito popanga RNA agarose gel electrophoresis.DNA yamitundu iwiri imakonda kuthamanga kwambiri mu TAE kuposa m'mabafa ena ndipo imatha kutopa panthawi yotalikirapo ya electrophoresis.Kuzungulira kwa buffer kapena kusintha kwa buffer panthawi yotalikirapo ya electrophoresis kumatha kuthana ndi kutsika kocheperako.Itha kugwiritsidwa ntchito mosiyanasiyana pophunzira kuyenda kwa DNA mu yankho.Popeza borate mu TBE buffer (Tris-Borate-EDTA Buffer, 10X Powder Pack, sc-296651) ndi inhibitor yamphamvu ya michere yambiri, TAE buffer imalimbikitsidwa poyang'ana ntchito za enzymatic za chitsanzo cha DNA.Mchere wa Tris acetate ndiwonso wotchinga wokhala ndi chidwi kwambiri pamayesero a ATP okhala ndi chiphamvu luciferase.

Kagwiritsidwe: TAE kuthamanga buffer ndiye malo omwe amagwiritsidwa ntchito kwambiri pa DNA agarose gel electrophoresis, ndipo amagwiritsidwanso ntchito pa RNA agarose gel electrophoresis.DNA yamitundu iwiri imakonda kuyenda mwachangu mu TAE kuposa m'mabafa ena, komanso imalephera kusambira chifukwa cha kuchepa kwa ma ion a buffer panthawi ya electrophoresis yayitali.Kuyendetsa njinga zamoto kapena kusinthana kwa buffer panthawi ya electrophoresis yayitali kumatha kubweza kuchuluka kwa buffer kutsika.2 Sungunulani buffer ya TAE yokhazikika kuti mupeze 1 mMTAE buffer yokhala ndi 40 mM Tris acetate ndi 1 mM EDTA, pH 8.3.1 mMTAE buffer itha kugwiritsidwa ntchito mu ma gels a agarose komanso ngati buffer.Pakuwongolera kwakukulu, tikulimbikitsidwa kuti magetsi ogwiritsidwa ntchito akhale otsika kuposa 5V/cm (mtunda pakati pa ma electrode a unit).

Kugwiritsa ntchito: TAE yoyendetsa buffer ndiye malo omwe amagwiritsidwa ntchito kwambiri pa DNA agarose gel electrophoresis mu Chemicalbook gel, ndipo amagwiritsidwanso ntchito popanga RNA agarose gel electrophoresis.DNA yamitundu iwiri imakonda kuyenda mwachangu mu TAE kuposa m'mabafa ena, komanso imalephera kusambira chifukwa cha kuchepa kwa ma ion a buffer panthawi ya electrophoresis yayitali.Kuyendetsa njinga zamoto kapena kusinthana kwa buffer panthawi ya electrophoresis yayitali kumatha kubweza kuchuluka kwa buffer kutsika.Buffer ya TAE yokhazikika idachepetsedwa kuti ipeze 1 mMTAE buffer yokhala ndi 40 mM Tris acetate ndi 1 mM EDTA, pH 8.3.1 mMTAE buffer itha kugwiritsidwa ntchito mu ma gels a agarose komanso ngati buffer.Pakuwongolera kwakukulu, tikulimbikitsidwa kuti magetsi ogwiritsidwa ntchito akhale otsika kuposa 5V/cm (mtunda pakati pa ma electrode a unit).

Ntchito: Pozindikira ATP ndi ziphaniphani luciferase, mankhwalawa ndiye chitetezo chovuta kwambiri;kuzindikira kwa glutamate.

Kumanga kosasunthika kwa [3H]l-glutamic acid kuzinthu zosalandira.

[3H]L-glutamic acid yomangirira ku machubu a microfuge ndi galasi adafufuzidwa m'mabafa anayi.Kumangirira zakumbuyo kuzinthu izi kunali kocheperako, koma kudakulitsidwa ndi kulowetsa kapena kuyamwa mu Tris-HCl ndi Tris-citrate buffer.Kumanga uku kunali kochepa kwambiri kapena Pamene HEPES-KOH, kapena Tris-acetate buffer idagwiritsidwa ntchito m'malo mwake.[3H] Kumanga kwa L-glutamate kumachubu a microfuge kunaletsedwa ndi L- koma osati D-isomers ya glutamate ndi aspartate.DL-2-amino-7-phosphonoheptanoic asidi nayenso sanalepheretse kumanga.Mankhwala ena omwe amawonetsa kuletsa kwapang'onopang'ono anali: N-methyl-D-aspartate, quisqualate, L-glutamic acid diethyl ester, N-methyl-L-aspartate, kainate, ndi 2-amino-4 -phosphonobutyrate.Kumanga kunaletsedwa ndi nembanemba za ubongo wa makoswe.Kumangirira kodalira mapuloteni [3H] glutamate kunapezedwa ndi kukonzekera kwa nembanemba kowumitsidwa mobwerezabwereza pamene kumangirira kunachitika mu Tris-acetate buffer.Ndikofunikira kuti Tris-acetate kapena HEPES -KOH buffer iyenera kugwiritsidwa ntchito mu glutamate bin ding assay.Ngati Tris-HCl kapena Tris-citrate buffer agwiritsidwa ntchito, kuyesa koyenera kuyenera kuchitidwa kuti kumangiridwe kumachubu a microfuge kapena zosefera zamagalasi.