1. Ho arola
Gel electrophoresis e arotsoe ka mefuta e tsepameng (ho kenyeletsoa li-gels tsa kholomo le li-gels tsa slab) le mefuta e rapameng (haholo-holo li-gels tsa slab) (Figure 6-18). Ka kakaretso, karohano e otlolohileng e phahame hanyenyane ho e holimo, empa ho lokisoa ha li-gel ho na le bonyane melemo e mene: ho na le ts'ehetso ka tlas'a gel eohle, e lumellang ho sebelisoa ha agarose e tlaase ea concentration; hoa khoneha ho lokisa lipoleiti tsa gel tsa agarose tsa litlhaloso tse fapaneng; ho lokisoa ha li-gel le ho kenya sampole ho bonolo haholoanyane; kamore ea electrophoresis e bonolo ho e haha le e theko e tlaase. Kaha electrophoresis e otlolohileng e etsoa ka poleiti ea gel ea agarose e qoeletseng ka ho feletseng hoo e ka bang 1mm ka tlas'a bokaholimo ba "electrophoresis buffer", e boetse e bitsoa "submerged electrophoresis".
Electrophoresis Tank DYCP-31DN bakeng sa agarose gel electrophoresis
Sistimi ea 2.Buffer
Karohanong ea nucleic acid, litsamaiso tse ngata li nka litsamaiso tse tsoelang pele. Li-buffer tsa electrophoresis tse sebelisoang ka tloaelo li kenyelletsa TBE (0.08mol/L Tris·HCl, pH 8.5, 0.08mol/L boric acid, 0.0024mol/L EDTA) buffer le THE (0.04mol/L Tris·HCl, pH 7.8, 0.02mol/L) sodium acetate, 0.0018mol/L EDTA) buffer. Li-buffers tsena ka kakaretso li lokisoa e le litharollo tsa stock tsa 10x 'me li hlapolotsoe ka bongata bo hlokahalang ha li sebelisoa. Litekanyetso tsa ho falla ha DNA ea mola le selikalikoe ka jeleng ea agarose li fapana ho latela buffer e sebelisitsoeng. Ho buffer, sekhahla sa phalliso ea DNA ea mola o moholo ho feta DNA e chitja, ha TBE buffer, se fapaneng ke 'nete.
3.Ho lokisetsa Gel ea Agarose
(1) Tokisetso ea Horizontal Agarose Gel
(a) Lokisetsa motsoako o hlokahalang oa gel ea agarose o sebelisa 1x electrophoresis buffer.
(b) Chesa agarose hore e qetelle e qhibiliha, e ka ba ka bateng ea metsi a belang, holim'a mochini o futhumatsang oa makenete, kapa ka microwave. Fokotsa tharollo ea agarose ho 55 ° C 'me u kenye dae ea ethidium bromide (EB) ho motsoako oa ho qetela oa 0.5 μg/ml.
(c) Koahela mathōko a likhalase kapa lipoleiti tsa acrylic ka gel e nyenyane ea agarose, eketsa kama, 'me u behe meno a khekhe ka 0.5 ~ 1.0 mm ka holim'a poleiti.
(d) Tšela tharollo ea gel ea agarose e qhibilihisitsoeng ka ho tsoelang pele ka khalase kapa hlobo ea acrylic plate (botenya bo itšetlehile ka boholo ba sampuli ea DNA), ho qoba ho kenngoa ha li-bubble tsa moea. E lumelle hore e tiee ka tlhaho mocheso oa kamore.
(e) Tlosa kama ka hloko ka mor'a ho tiisa ka ho feletseng. Eketsa tekanyo e loketseng ea "electrophoresis buffer" tanka ea gel, ho etsa bonnete ba hore poleiti ea gel e qoelisoa hoo e ka bang 1 mm ka tlas'a bokaholimo ba "electrophoresis buffer".
(2) Tokiso ea Gel ea Vertical Agarose
(a) Tlosa mafura kapa masalla lipoleiting tsa khalase ka ho hlatsoa ka ethanol.
(b) Beha lipolanete tsa spacer lipakeng tsa matamo a ka pele le a ka morao, hokahanya mathōko a li-spacer plates le matamo a ka pele le a ka morao, 'me u a tiise ka li-clamp.
(c) Kenya 2% agarose ka har'a 1x buffer lipakeng tsa mephetho ea lipoleiti tsa spacer ho etsa polaka e phahameng ea agarose ea 1 cm tlase ho phaposi ea ho akhela gel.
(d) Tšela gel ea agarose e qhibilihisitsoeng sebakeng se lakatsehang, se lokiselitsoeng ka 1x buffer, ka kamoreng ea gel ho fihlela ho 1 cm ka tlase holimo.
(e) Kenya kama, ho qoba ho tšoasa lipululana tsa moea tlas'a meno a khekhe. Ka linako tse ling, wrinkles ka 'na hlaha ka khekhe meno nakong agarose pholileng gel; maemong a joalo, eketsa agarose e qhibilihisitsoeng holimo ho e tiisa.
(f) Tlosa kama. Ho thibela ho dutla ha buffer sekoting sa ho palamisa, tiisa khokahano lipakeng tsa poleiti ea gel ea agarose le phaposi ea electrophoresis ka 2% ea agarose mme o kenye palo e hlokahalang ea buffer.
(g) Eketsa 1x electrophoresis buffer kamoreng ea gel.
(h) Kenya lisampole tsa DNA ka hloko holim'a gel ea agarose ka tlas'a buffer.
Boitsebiso bo eketsehileng mabapi le tsebo ea motheo ka agarose gel electrophoresis, re tla arolelana bekeng e tlang. Ke lakatsa eka lintlha tsena li ka thusa tekong ea hau.
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Nako ea poso: Dec-07-2023