Njira Zoyambira za Agarose Gel Electrophoresis(2)

Chitsanzo Kukonzekera ndiKutsegula

Chifukwa chakugwiritsa ntchito kachitidwe ka buffer mosalekeza popanda kuyika gel nthawi zambiri, zitsanzo ziyenera kukhala ndi ndende yoyenera komanso voliyumu yaying'ono. Gwiritsani ntchito apipettekuti muwonjezere pang'onopang'ono chitsanzo, ndi 5-10 μg pachitsime, kuti mupewe kuchepa kwakukulu kwa chisankho. Litipotsegula chitsanzo, magetsi ayenera kuzimitsidwa. Chitsanzocho chiyenera kukhala ndi utoto wosonyeza (0.025% bromophenol buluu kapena lalanje) ndi sucrose (10-15%) kapena glycerol (5-10%) kuti awonjezere kachulukidwe, kuyika kwambiri chitsanzo, ndi kupewa kufalikira. Komabe, nthawi zina sucrose kapena glycerol imatha kuyambitsa magulu ooneka ngati U muzotsatira za electrophoresis, zomwe zitha kupewedwa pogwiritsa ntchito 2.5% Ficoll (polyvinylpyrrolidone).

Electrophoresis

Magetsi a electrophoresis ndi 5-15 V / cm, nthawi zambiri pafupifupi 10 V / cm. Pakulekanitsa mamolekyu akuluakulu, magetsi ayenera kukhala otsika, nthawi zambiri osapitirira 5 V / cm.

Kudetsa

Fluorescent dye ethidium bromide (EB) imagwiritsidwa ntchito kwambiri podetsa poyang'ana magulu a DNA mu gel agarose. EB imatha kuyika pakati pa magawo awiri a mamolekyu a DNA, kupangitsa EB kumangika ndi DNA. Kuyamwa kwa 260 nm ultraviolet kuwala ndi DNA kumasamutsidwa ku EB, ndipo EB yomangidwa imatulutsa fluorescence pa 590 nm m'chigawo chofiira-lalanje cha kuwala kowonekera. Kuthira gel mu njira ya 1 mmol/L MgSO4 kwa ola limodzi kumatha kuchepetsa fluorescence yakumbuyo chifukwa cha EB yosamangidwa, zomwe zimathandizira kuzindikira kachulukidwe kakang'ono ka DNA.

Utoto wa EB uli ndi maubwino angapo: ndi wosavuta kugwiritsa ntchito, suphwanya ma nucleic acid, umakhala ndi chidwi kwambiri, ndipo ukhoza kuyipitsa DNA ndi RNA. EB ikhoza kuwonjezeredwa ku zitsanzo ndikutsatiridwa pogwiritsa ntchito kuyamwa kwa UV nthawi iliyonse. Pambuyo pakudetsa, EB imatha kuchotsedwa ndikuchotsa ndi n-butanol.

Komabe, utoto wa EB ndi mutagen wamphamvu, ndipo kusamala, monga kuvala magolovesi a polyethylene, kuyenera kutengedwa pogwira. Acridine lalanje ndi utoto womwe umagwiritsidwa ntchito nthawi zambiri chifukwa umatha kusiyanitsa pakati pa nucleic acids (DNA, RNA) yokhala ndi chingwe chimodzi kapena iwiri. Imawonetsa ma fluorescence obiriwira (530 nm) a nucleic acid okhala ndi mizere iwiri ndi fluorescence yofiira (640 nm) ya nucleic acid yokhala ndi chingwe chimodzi. Kuphatikiza apo, utoto wina monga methylene blue, methylene wobiriwira, ndi quinoline B ungagwiritsidwe ntchito podetsa.

Beijing Liuyi Biotechnology Co. Ltd (Liuyi Biotechnology) yakhala ikupanga zida za electrophoresis kwa zaka zopitilira 50 ndi gulu lathu laukadaulo laukadaulo ndi R&D pakati. Tili ndi mzere wodalirika komanso wathunthu wopanga kuchokera pamapangidwe mpaka kukayendera, ndi nyumba yosungiramo zinthu, komanso chithandizo chamalonda. Zogulitsa zathu zazikulu ndi Electrophoresis Cell (thanki / chipinda), Electrophoresis Power Supply, Blue LED Transilluminator, UV Transilluminator, Gel Image & Analysis System etc.

Tsopano tikuyang'ana mabwenzi, onse a OEM electrophoresis thanki ndi ogulitsa amalandiridwa.

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