Muenzaniso Kugadzirira uyeLoading
Nekuda kwekushandiswa kweinoenderera mberi buffer system isina kurongedza gel muzviitiko zvakawanda, masampuli anofanirwa kuve neakakodzera kusungwa uye vhoriyamu diki. Shandisa apipettekuwedzera zvishoma nezvishoma sampu, ine 5-10 μg patsime, kudzivirira kuderera kukuru mukugadziriswa. Rinikurodha muenzaniso, magetsi anofanira kuvharwa. Sample inofanira kunge iine dhayi yechiratidzo (0.025% bromophenol bhuruu kana orenji) uye sucrose (10-15%) kana glycerol (5-10%) kuti iwedzere kuwanda kwayo, kutarisisa sampuli, uye kudzivirira kupararira. Zvisinei, dzimwe nguva sucrose kana glycerol inogona kukonzera mabhandi akafanana neU mumigumisiro ye electrophoresis, iyo inogona kudzivirirwa nekushandisa 2.5% Ficoll (polyvinylpyrrolidone).
Electrophoresis
Iyo voltage ye electrophoresis ndeye 5-15 V/cm, kazhinji inotenderera 10 V/cm. Nekuparadzaniswa kwemamorekuru makuru, voltage inofanira kunge yakaderera, kazhinji isingapfuuri 5 V / cm.
Staining
Fluorescent dye ethidium bromide (EB) inowanzoshandiswa kusvibisa kutarisa mabhendi eDNA muagarose gel. EB inogona kuisa pakati pemabhesi maviri emamorekuru eDNA, zvichiita kuti EB ibatane neDNA. Kutorwa kwe260 nm ultraviolet chiedza neDNA kunoendeswa kuEB, uye EB yakasungwa inobudisa fluorescence pa 590 nm munharaunda tsvuku-orenji yechiedza chinooneka. Kusvibisa gel mu 1 mmol / L MgSO4 mhinduro kweawa imwe kunogona kuderedza kumashure fluorescence inokonzerwa neB isina kuvharwa, zvichiita kuti kuonekwa kwezvishoma zvishoma zveDNA.
EB dhayi ine zvakati wandei zvakanakira: iri nyore kushandisa, haityori nucleic acids, ine yakanyanya senitivity, uye inogona kusvibisa zvese DNA neRNA. EB inogona kuwedzerwa kumuenzaniso uye kuteverwa uchishandisa UV kunyura chero nguva. Mushure mekusvibisa, EB inogona kubviswa nekubvisa ne n-butanol.
Zvisinei, EB dhayi ine simba mutagen, uye zvidziviriro, zvakadai sokupfeka magirovhosi epolyethylene, zvinofanira kutorwa panguva yekubata. Acridine orenji zvakare dhayi inowanzoshandiswa nekuti inogona kusiyanisa pakati peiyo-yakasungwa uye kaviri-yakasungwa nucleic acids (DNA, RNA). Inoratidza green fluorescence (530 nm) yekaviri-stranded nucleic acids uye tsvuku fluorescence (640 nm) ye single-stranded nucleic acids. Pamusoro pezvo, mamwe madhayi akadai semethylene bhuruu, methylene girini, uye quinoline B anogona kushandiswa kusvibisa.
Beijing Liuyi Biotechnology Co. Ltd (Liuyi Biotechnology) ine hunyanzvi mukugadzira zviridzwa zve electrophoresis kweanopfuura makore makumi mashanu nechikwata chedu chehunyanzvi chehunyanzvi uye R&D centre. Isu tine yakavimbika uye yakakwana yekugadzira mutsara kubva padhizaini kusvika pakuongorora, uye imba yekuchengetera, pamwe nerutsigiro rwekushambadzira. Zvigadzirwa zvedu zvikuru ndeElectrophoresis Cell (tangi/chamber), Electrophoresis Power Supply, Blue LED Transilluminator, UV Transilluminator, Gel Image & Analysis System etc.
Isu tave kutsvaga vanobatika, vese OEM electrophoresis tank uye vanogovera vanogamuchirwa.
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Nguva yekutumira: Zvita-20-2023