Njira Zoyambira za Agarose Gel Electrophoresis(1)

1. Gulu

Gel electrophoresis lagawidwa mu mitundu ofukula (kuphatikizapo gel osakaniza ndime ndi gel osakaniza slab) ndi mitundu yopingasa (makamaka slab gel osakaniza) (Chithunzi 6-18). Kawirikawiri, kupatukana kowongoka kumakhala kopambana pang'ono kuposa kopingasa, koma kukonzekera gel osakaniza kumakhala ndi ubwino zinayi: pali chithandizo pansi pa gel osakaniza, kulola kugwiritsa ntchito agarose otsika kwambiri; ndizotheka kukonzekera mbale za gel agarose zamitundu yosiyanasiyana; Kukonzekera kwa gel ndi kuyika zitsanzo ndizosavuta; chipinda cha electrophoresis ndi chosavuta kupanga komanso chotsika mtengo. Popeza electrophoresis yopingasa imachitidwa ndi mbale ya agarose gel yomizidwa kwathunthu pafupifupi 1mm pansi pamadzi a electrophoresis buffer, imatchedwanso submerged electrophoresis.

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Electrophoresis Tank DYCP-31DN ya agarose gel electrophoresis

2.Buffer System

Mu kupatukana kwa nucleic acid, machitidwe ambiri amatenga machitidwe opitilira. Ma buffers a electrophoresis omwe amagwiritsidwa ntchito kwambiri ndi TBE (0.08mol/L Tris·HCl, pH 8.5, 0.08mol/L boric acid, 0.0024mol/L EDTA) buffer ndi THE (0.04mol/L Tris·HCl, pH 7.8, 0.02mol/L) sodium acetate, 0.0018mol/L EDTA) buffer. Ma buffers awa nthawi zambiri amakonzedwa ngati 10x stock solutions ndipo amachepetsedwa mpaka ndende yofunikira akagwiritsidwa ntchito. Kusamuka kwa DNA yozungulira ndi yozungulira mu gel agarose imasiyana ndi buffer yomwe imagwiritsidwa ntchito. Mu buffer, kuchuluka kwa kusamuka kwa DNA yozungulira ndikokulirapo kuposa DNA yozungulira, pomwe mu TBE buffer, zosiyana ndizowona.

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3.Kukonzekera kwa Agarose Gel

(1) Kukonzekera kwa Horizontal Agarose Gel

(a) Konzani kuchuluka kwa gel osakaniza agarose pogwiritsa ntchito 1x electrophoresis buffer.

(b) Kutenthetsa agarose kuti amalize kusungunuka m'madzi osamba owiritsa, pamagetsi osonkhezera maginito, kapena mu microwave. Muziziziritsa njira ya agarose mpaka 55°C ndipo onjezerani utoto wa ethidium bromide (EB) mpaka kufika pa 0.5 μg/ml.

(c) Tsekani m'mphepete mwa galasi kapena mbale za acrylic ndi gel osakaniza pang'ono, onjezani chisa, ndikuyika mano achisa pafupifupi 0.5 ~ 1.0 mm pamwamba pa mbaleyo.

(d) Thirani njira yosungunuka ya agarose gel mosalekeza mu galasi kapena nkhungu ya acrylic plate (kuchuluka kumadalira kuchuluka kwa zitsanzo za DNA), kupewa kuyambitsa thovu la mpweya. Lolani kuti likhale lolimba mwachibadwa kutentha kwapakati.

(e) Chotsani chisacho mosamala mukachilimbitsa. Onjezani kuchuluka koyenera kwa bafa ya electrophoresis ku tanki ya gel, kuwonetsetsa kuti mbale ya gel imamizidwa pafupifupi 1 mm pansi pa chotchinga cha electrophoresis.

(2) Kukonzekera kwa Vertical Agarose Gel

(a) Chotsani mafuta kapena zotsalira mu mbale zagalasi posamba ndi ethanol.

(b) Ikani mbale za spacer pakati pa madamu akutsogolo ndi akumbuyo, gwirizanitsani m'mphepete mwa madamu akutsogolo ndi akumbuyo, ndipo muwateteze ndi zingwe.

(c) Onjezani 2% agarose mu 1x bafa pakati pa m'mphepete mwa mbale za spacer kuti mupange pulagi ya agarose yotalika 1 cm pansi pa chipinda choponyera gel.

(d) Thirani gel osungunuka agarose pamalo omwe mukufuna, okonzedwa mu 1x buffer, muchipinda cha gel mpaka 1 cm pansi pamwamba.

(e) Ikani chisa, kupewa kutchera thovu la mpweya pansi pa mano achisa. Nthawi zina, makwinya angawonekere pachisa mano pa kuzirala kwa gel osakaniza agarose; Zikatero, ingowonjezerani agarose wosungunuka pamwamba kuti alimbitse.

(f) Chotsani chisa. Kuti mupewe kutayikira kwa bafa potsegula, sindikizani kulumikizana pakati pa mbale ya agarose gel ndi chipinda cha electrophoresis ndi 2% agarose ndikuwonjezera kuchuluka kofunikira.

(g) Onjezani 1x electrophoresis buffer kuchipinda cha gel.

(h) Lowetsani mosamala zitsanzo za DNA pa gel agarose pansi pa buffer.

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Zambiri zokhudzana ndi chidziwitso choyambirira cha agarose gel electrophoresis, tidzagawana nawo sabata yamawa. Ndikukhumba kuti chidziwitsochi chikhale chothandizira kuyesa kwanu.

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.

Ngati muli ndi pulani yogulira zinthu zathu, chonde musazengereze kulumikizana nafe. Mutha kutitumizira uthenga pa imelo[imelo yotetezedwa]kapena[imelo yotetezedwa], kapena chonde tiyimbireni ku +86 15810650221 kapena onjezani Whatsapp +86 15810650221, kapena Wechat: 15810650221.

Chonde Jambulani nambala ya QR kuti muwonjezere pa WhatsApp kapena WeChat.

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Nthawi yotumiza: Dec-07-2023