Amasu Ayisisekelo e-Agarose Gel Electrophoresis(2)

Ukulungiselela Isampula kanyeIyalayisha

Ngenxa yokusetshenziswa kwesistimu ye-buffer eqhubekayo ngaphandle kokupakisha ijeli ezimweni eziningi, amasampula kufanele abe nokugxila okufanele kanye nevolumu encane. Sebenzisa ai-pipetteukwengeza kancane isampula, ngo-5-10 μg emthonjeni ngamunye, ukugwema ukwehla okuphawulekayo kokuxazulula. Niniilayisha isampula, ukunikezwa kwamandla kumele kucishwe. Isampula kufanele iqukathe udayi wenkomba (0.025% we-bromophenol oluhlaza okwesibhakabhaka noma owolintshi) kanye ne-sucrose (10-15%) noma i-glycerol (5-10%) ukuze kukhuliswe ukuminyana kwayo, kugxilise isampula, futhi kuvimbele ukusakazeka. Nokho, ngezinye izikhathi i-sucrose noma i-glycerol ingase ibangele amabhande amise okwe-U emiphumeleni ye-electrophoresis, engagwenywa ngokusebenzisa i-2.5% Ficoll (polyvinylpyrrolidone).

I-Electrophoresis

I-voltage ye-electrophoresis ingu-5-15 V/cm, imvamisa ibe ngu-10 V/cm. Ukuze kuhlukaniswe ama-molecule amakhulu, i-voltage kufanele ibe ngaphansi, ngokuvamile ingabi ngaphezu kwe-5 V / cm.

Ukufaka amabala

Udayi we-fluorescent ethidium bromide (EB) uvame ukusetshenziselwa ukungcolisa ukuze ubheke amabhande e-DNA kujeli ye-agarose. I-EB ingafaka phakathi kwamapheya esisekelo ama-molecule e-DNA, okubangele ukuthi i-EB ihlangane ne-DNA. Ukumuncwa kokukhanya kwe-ultraviolet okungu-260 nm nge-DNA kudluliselwa ku-EB, futhi i-EB eboshiwe ikhipha i-fluorescence ku-590 nm endaweni ebomvu-owolintshi ye-spectrum yokukhanya ebonakalayo. Ukungcolisa ijeli kusixazululo esingu-1 mmol/L MgSO4 ihora eli-1 kunganciphisa i-fluorescence yangemuva ebangelwa i-EB engaboshiwe, kube lula ukutholwa kwenani elincane le-DNA.

Idayi ye-EB inezinzuzo eziningana: kulula ukuyisebenzisa, ayiphuli ama-nucleic acid, inokuzwela okuphezulu, futhi ingangcolisa kokubili i-DNA ne-RNA. I-EB ingangezwa kusampula futhi ilandelelwe kusetshenziswa ukumuncwa kwe-UV noma kunini. Ngemuva kokugcoba, i-EB ingasuswa ngokukhipha nge-n-butanol.

Kodwa-ke, udayi we-EB uyi-mutagen enamandla, futhi izinyathelo zokuphepha, njengokugqoka amagilavu ​​e-polyethylene, kufanele zithathwe ngesikhathi sokuphatha. I-Acridine orange nayo ingudayi osetshenziswa kakhulu ngoba ingakwazi ukuhlukanisa phakathi kwama-nucleic acid anemicu eyodwa naphindwe kabili (DNA, RNA). Ibonisa i-fluorescence eluhlaza (530 nm) yama-nucleic acid anemicu ephindwe kabili kanye ne-fluorescence ebomvu (640 nm) yama-nucleic acid anemigqa eyodwa. Ukwengeza, abanye odayi abanjenge-methylene blue, i-methylene eluhlaza, kanye ne-quinoline B bangasetshenziselwa ukugcoba.

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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