Amasu Ayisisekelo e-Agarose Gel Electrophoresis(1)

1. Ukwahlukanisa

I-Gel electrophoresis ihlukaniswe yaba izinhlobo eziqondile (kuhlanganise namajeli ekholomu namajeli e-slab) nezinhlobo ezivundlile (ikakhulukazi ama-slab gels) (Umfanekiso 6-18). Ngokuvamile, ukuhlukaniswa okuqondile kuphakeme kancane kunezingqimba, kodwa ukulungiswa kwejeli enezingqimba kunezinzuzo okungenani ezine: kukhona ukusekelwa ngaphansi kwayo yonke ijeli, okuvumela ukusetshenziswa kwe-agarose ephansi yokugxila; kungenzeka ukulungisa amapuleti ejeli e-agarose emininingwane ehlukene; ukulungiswa kwejeli nokulayisha isampula kulula kakhulu; Igumbi le-electrophoresis kulula ukulakha futhi lingabizi. Njengoba i-electrophoresis evundlile yenziwa ngepuleti yejeli ye-agarose ecwiliswe ngokuphelele cishe ngo-1mm ngaphansi kobuso be-electrophoresis buffer, ibizwa nangokuthi i-submerged electrophoresis.

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I-Electrophoresis Tank DYCP-31DN ye-agarose gel electrophoresis

2.Isistimu yebhafa

Ekuhlukaniseni i-nucleic acid, amasistimu amaningi athatha amasistimu aqhubekayo. Amabhafa we-electrophoresis asetshenziswa kakhulu afaka i-TBE (0.08mol/L Tris·HCl, pH 8.5, 0.08mol/L boric acid, 0.0024mol/L EDTA) buffer kanye ne-THE (0.04mol/L Tris·HCl, pH 7.8, 0.02mol/L) i-acetate ye-sodium, i-0.0018mol/L EDTA) ibhafa. Lawa mabhafa ngokuvamile alungiswa njengezixazululo zesitoko eziyi-10x futhi ahlanjululwe ekugxiliseni okudingekayo lapho esetshenziswa. Amazinga okufuduka e-DNA yomugqa neyindilinga kujeli ye-agarose ayahluka ngesigcinalwazi esisetshenzisiwe. KU-BUffer, izinga lokufuduka kwe-DNA yomugqa likhulu kunele-DNA eyindilinga, kuyilapho ku-TBE buffer, okuphambene kuyiqiniso.

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3.Ukulungiswa kwe-Agarose Gel

(1) Ukulungiswa Kwejeli ye-Agarose Evundlile

(a) Lungiselela ukuhlangana okudingekile kwejeli ye-agarose usebenzisa i-1x electrophoresis buffer.

(b) Shisisa i-agarose ukuze uqedele ukuncibilika kungaba kubhavu wamanzi abilayo, kumshini ozibuthe oshukumisayo, noma kuhhavini we-microwave. Pholisa isisombululo se-agarose sibe ngu-55°C bese wengeza udayi we-ethidium bromide (EB) ekuhlanganiseni kokugcina kuka-0.5 μg/ml.

(c) Vala amaphethelo engilazi noma amapuleti e-acrylic ngenani elincane lejeli ye-agarose, engeza ikamu, bese ubeka amazinyo ekama cishe ngo-0.5~1.0 mm ngaphezu kwepuleti.

(d) Thela isisombululo sejeli ye-agarose encibilikisiwe ngokuqhubekayo engilazini noma esikhunjeni sepuleti le-acrylic (ugqinsi luncike kumthamo wesampula ye-DNA), ugweme ukwethulwa kwamabhamuza omoya. Ivumele iqine ngokwemvelo ekamelweni lokushisa.

(e) Khipha ngokucophelela ikama ngemva kokuqina okuphelele. Engeza inani elifanele le-electrophoresis buffer ethangini lejeli, uqinisekise ukuthi ipuleti lejeli licwiliswa emanzini cishe ngo-1 mm ngaphansi kobuso be-electrophoresis buffer.

(2) Ukulungiswa Kwejeli ye-Agarose Emile

(a) Khipha amafutha noma izinsalela ezitsheni zengilazi ngokugeza nge-ethanol.

(b) Beka amapuleti asemkhathini phakathi kwamadamu angaphambili nangemuva, qondanisa amaphethelo amapuleti e-spacer namadamu angaphambili nangemuva, futhi uwaqinise ngezinsimbi.

(c) Engeza i-agarose engu-2% kubhafa engu-1x phakathi kwamaphethelo amapuleti e-spacer ukuze wenze ipulaki ye-agarose ephakeme engu-1 cm ngaphansi kwegumbi lokuphonsa ijeli.

(d) Thela ijeli ye-agarose encibilikisiwe endaweni oyifunayo, elungiselelwe ku-1x buffer, ekamelweni lejeli kuze kufike ku-1 cm ngaphansi phezulu.

(e) Faka ikama, ugweme ukubamba amabhamuza omoya ngaphansi kwamazinyo ekama. Ngezinye izikhathi, imibimbi ingase ivele emazinyweni ekamu ngesikhathi sokupholisa ijeli ye-agarose; ezimweni ezinjalo, mane ungeze i-agarose encibilikisiwe phezulu ukuze uyiqinise.

(f) Khipha ikama. Ukuze uvimbele ukuvuza kwebhafa endaweni yokulayisha, vala uxhumano phakathi kwepuleti lejeli le-agarose negumbi le-electrophoresis nge-agarose engu-2% bese wengeza inani elidingekayo le-buffer.

(g) Engeza i-1x electrophoresis buffer ekamelweni lejeli.

(h) Layisha ngokucophelela amasampula e-DNA kujeli ye-agarose ngaphansi kwesigcinalwazi.

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Ulwazi oluthe xaxa mayelana nolwazi oluyisisekelo mayelana ne-agarose gel electrophoresis, sizokwabelana ngesonto elizayo. Fisa ukuthi lolu lwazi lube usizo ekuhloleni kwakho.

I-Beijing Liuyi Biotechnology Co. Ltd (i-Liuyi Biotechnology) ikhethekile ekukhiqizeni amathuluzi e-electrophoresis iminyaka engaphezu kwengu-50 neqembu lethu lezobuchwepheshe eliqeqeshiwe kanye nesikhungo se-R&D. Sinomugqa wokukhiqiza onokwethenjelwa nophelele kusukela ekuklanyweni kuya ekuhlolweni, kanye nendawo yokugcina impahla, kanye nokwesekwa kokumaketha. Imikhiqizo yethu eyinhloko i-Electrophoresis Cell (ithangi/igumbi), i-Electrophoresis Power Supply, i-Blue LED Transilluminator, i-UV Transilluminator, iGel Image & Analysis System njll.

Manje sifuna ozakwethu, kokubili ithangi le-OEM electrophoresis kanye nabasabalalisi bamukelekile.

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Isikhathi sokuthumela: Dec-07-2023