Ubuchwephesha obusisiseko be-Agarose Gel Electrophoresis(1)

1. Ukuhlelwa

I-Gel electrophoresis yahlulwe yaba ziindidi ezithe nkqo (kubandakanywa iigels zekholamu kunye neegels ze-slab) kunye neentlobo ezithe tye (ikakhulukazi i-slab gels) (Umfanekiso 6-18). Ngokuqhelekileyo, ukuhlukana okuthe tye kuphezulu kancinane kune-horizontal, kodwa ukulungiswa kwe-gel ene-horizontal ineenzuzo ezine ubuncinane: kukho inkxaso phantsi kwe-gel yonke, evumela ukusetyenziswa kwe-agarose ephantsi-concentration; kunokwenzeka ukulungiselela iipleyiti zejeli ze-agarose zeempawu ezahlukeneyo; ukulungiswa kwejeli kunye nokulayisha isampuli kulula ngakumbi; igumbi le-electrophoresis kulula ukulakha kunye neendleko. Ekubeni i-electrophoresis ethe tyaba yenziwa ngepleyiti yejeli ye-agarose entywiliselwe ngokupheleleyo malunga ne-1mm ngaphantsi komphezulu we-electrophoresis buffer, ikwabizwa ngokuba yi-submerged electrophoresis.

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

2.Isistim yeBuffer

Ekuhlukaneni kwe-nucleic acid, ezininzi iinkqubo zisebenzisa iinkqubo eziqhubekayo. Izithinteli ze-electrophoresis ezisetyenziswa ngokuqhelekileyo ziquka i-TBE (0.08mol/L Tris·HCl, pH 8.5, 0.08mol/L boric acid, 0.0024mol/L EDTA) buffer kunye THE (0.04mol/L Tris·HCl, pH 7.8, 0.02mol/L) i-acetate ye-sodium, i-0.0018mol / L EDTA) i-buffer. Ezi buffers zilungiswa ngokubanzi njengezisombululo zesitokhwe ezili-10x kwaye zixutywe kugxininiso olufunekayo xa zisetyenziswa. Amazinga okufuduka kwe-DNA yomgca kunye nesetyhula kwijeli ye-agarose iyahluka kunye nesithinteli esisetyenzisiweyo. Kwi-buffer, izinga lokufuduka kwe-linear DNA likhulu kunelo le-DNA ejikelezayo, ngelixa kwi-TBE buffer, okwahlukileyo kuyinyani.

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3.Ukulungiswa kweGel yeAgarose

(1) Ukulungiswa kwe-Horizontal Agarose Gel

(a) Lungiselela i-concentration efunekayo yejeli ye-agarose usebenzisa i-1x electrophoresis buffer.

(b) Fudumeza iagarose ukugqibezela ukunyibilika nokuba kukwibhafu yamanzi abilayo, kwisivunguvungu sikamagnetic, okanye kwimicrowave. Pholisa isisombululo se-agarose kwi-55 ° C kwaye ungeze idayi ye-ethidium bromide (EB) kwi-concentration yokugqibela ye-0.5 μg / ml.

(c) Vala imiphetho yeglasi okanye iipleyiti zeacrylic ngomlinganiselo omncinane wejeli yeagarose, yongeza ikama, uze ubeke amazinyo ekama malunga ne-0.5 ~ 1.0 mm ngaphezu kwepleyiti.

(d) Galela isisombululo sejeli ye-agarose enyibilikisiweyo ngokuqhubekayo kwiglasi okanye i-acrylic plate mold (ubukhulu buxhomekeke kumthamo wesampuli ye-DNA), ukuphepha ukuqaliswa kwamaqamza omoya. Mayiqinise ngokwemvelo kwiqondo lokushisa.

(e) Yisuse ngononophelo ikama emva kokuqina ngokupheleleyo. Yongeza umlinganiselo ofanelekileyo we-electrophoresis buffer kwitanki yejeli, uqinisekise ukuba ipleyiti yejeli ifakwe emanzini malunga ne-1 mm ngaphantsi komphezulu we-electrophoresis buffer.

(2) Ukulungiswa kweGel ye-Agarose ye-Vertical

(a) Susa igrisi okanye intsalela kwiipleyiti zeglasi ngokuhlamba nge-ethanol.

(b) Beka iipleyiti ezisemajukujukwini phakathi kwamadama angaphambili nangasemva, lungelelanisa imiphetho yeepleyiti zesiphekepheke kunye namadama angaphambili nangasemva, kwaye uwaqinise ngezibophelelo.

(c) Yongeza i-2% yeagarose kwi-1x buffer phakathi kweencam zeepleyiti zesikhewu ukwenza iplagi ye-agarose eyi-1 cm ephezulu emazantsi egumbi lokujula ijeli.

(d) Galela ijeli ye-agarose enyibilikisiweyo kwindawo oyifunayo, elungiselelwe kwi-1x buffer, kwigumbi lejeli ukuya kwi-1 cm ngaphantsi phezulu.

(e) Faka ikama, ukunqanda ukubambisa amaqamza omoya phantsi kwamazinyo ekama. Ngamanye amaxesha, imibimbi inokuvela kumazinyo ekama ngexesha lokupholisa ijeli ye-agarose; kwiimeko ezinjalo, yongeza ngokulula iagarose enyibilikisiweyo phezulu ukuyiqinisa.

(f) Susa ikama. Ukuthintela ukuvuza kwe-buffer kwindawo yokulayishwa, vala umdibaniso phakathi kwepleyiti yejeli ye-agarose kunye negumbi le-electrophoresis nge-2% ye-agarose kwaye wongeze inani elifunekayo le-buffer.

(g) Yongeza i-1x electrophoresis buffer kwigumbi lejeli.

(h) Layisha ngononophelo iisampulu zeDNA kwijeli yeagarose ngaphantsi kwesithinteli.

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Olunye ulwazi malunga nolwazi olusisiseko malunga ne-agarose gel electrophoresis, siya kubelana ngeveki ezayo. Ndinqwenela ukuba olu lwazi lube luncedo kuvavanyo lwakho.

I-Beijing Liuyi Biotechnology Co. Ltd (i-Liuyi Biotechnology) ikhethekileyo ekuveliseni izixhobo ze-electrophoresis iminyaka engaphezu kwama-50 kunye neqela lethu lobuchwephesha lobuchwephesha kunye neziko le-R&D. Sinomgca wemveliso othembekileyo nopheleleyo ukusuka kuyilo ukuya ekuhlolweni, kunye nendawo yokugcina impahla, kunye nenkxaso yokuthengisa. Iimveliso zethu eziphambili ziyi-Electrophoresis Cell (itanki / igumbi), i-Electrophoresis Power Supply, i-Blue LED Transilluminator, i-UV Transilluminator, i-Gel Image & Analysis System njl.

Ngoku sikhangela amaqabane, zombini itanki ye-OEM electrophoresis kunye nabasasazi bamkelwe.

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Ixesha lokuposa: Dec-07-2023