Gel electrophorei ni bumwe muburyo bukomeye bukoreshwa muri biologiya ya molekuline yo gusesengura ADN. Ubu buryo bukubiyemo kwimuka kw'ibice bya ADN binyuze muri gel, aho bitandukanijwe ukurikije ubunini cyangwa imiterere. Ariko, waba warigeze uhura namakosa mugihe cya electrophoreis yawe, nka bande zasizwe kuri geli ya agarose, cyangwa ntamutwe uri kuri gel? Niki gishobora kuba nyirabayazana w'aya makosa?
Abatekinisiye bacu bavuze muri make abashakanye bakemura ibibazo hano kugirango ubone.
1. Amavuta asize kuri agarose gel
●ADN yarangiritse. Irinde kwanduza nuclease.
Buff Buffer ya electrophoreis ntabwo ari shyashya. Nyuma yo gukoresha inshuro nyinshi ya buffer ya electrophoreis, imbaraga za ionic ziragabanuka, kandi agaciro ka pH kiyongera, bityo ubushobozi bwa buffer bugabanuka, bikagira ingaruka kuri electrophorei. Birasabwa gusimbuza buffer ya electrophoreis kenshi.
Conditions Imiterere ya electrophoreis idakwiye yakoreshejwe. Ntukemere ko voltage irenga 20 V / cm, kandi ugumane ubushyuhe <30 ° C mugihe cya electrophorei. Kuri ADN nini ya electrophoreis, ubushyuhe bugomba kuba <15 ° C. Reba buffer ya electrophoreis ifite ubushobozi buhagije.
AD ADN nyinshi zipakiye kuri gel. Kugabanya ingano ya ADN.
Umunyu mwinshi muri ADN. Koresha imvura ya Ethanol kugirango ukureho umunyu urenze.
● ADN yari yanduye proteyine. Koresha ibiyikuramo bya fenol kugirango ukureho poroteyine yateye imbere.
AD ADN yamaganwe. Ntugashyuhe mbere ya electrophoreis. Koresha ADN muri buffer hamwe na 20 mM NaCl.
2. Anomalies kwimuka kwitsinda rya ADN
Kuvugurura urubuga rwa COS rwa λIcyiciro cya III. Shyushya ADN muminota 5 munsi ya 65 ° C mbere ya electrophoreis, hanyuma uyikonjeshe kuri ice ice muminota 5.
Conditions Imiterere ya electrophoreis idakwiye yakoreshejwe. Ntukemere ko voltage irenga 20 V / cm, kandi ugumane ubushyuhe <30 ° C mugihe cya electrophorei. Reba amashanyarazi ya electrophoreis afite ubushobozi buhagije bwa buffer.
AD ADN yamaganwe. Ntugashyuhe mbere ya electrophoreis. Koresha ADN muri buffer hamwe na 20 mM NaCl.
3. Kunanirwa cyangwa kutagira imirongo ya ADN kuri gel ya agarose
● Nta mubare uhagije cyangwa ubunini bwa ADN bwapakiye kuri gel. Ongera umubare wa ADN. Polyacrylamide gel electrophorei irumva gato kuruta agarose electrophoreis, kandi gupakira ibintu bishobora kugabanuka uko bikwiye.
AD ADN yangiritse. Irinde kwanduza nuclease.
ADN ADN yahawe amashanyarazi kuri gel. Electrophorese gel mugihe gito, koresha voltage yo hasi, cyangwa ukoreshe gel hejuru.
Source Inkomoko yumucyo idakwiye yakoreshejwe mugushushanya ADN ya etidium bromide. Koresha uburebure buke (254 nm) W urumuri kugirango urusheho kwiyumvisha ibintu.
4. Amatsinda ya ADN yabuze
●Ingano ntoya ADN yashizwemo amashanyarazi kuri gel. Electrophorese gel mugihe gito, koresha voltage yo hasi, cyangwa ukoreshe gel hejuru.
Biragoye gutandukanya imirongo ya ADN ya molekile isa. Ongera igihe cya electrophoreis, hanyuma urebe kwibandaya gel kugirango umenye neza gel ijanisha gukoreshwa.
AD ADN yamaganwe. Ntugashyuhe mbere ya electrophoreis. Koresha ADN muri buffer hamwe na 20 mM NaCl.
Ands Imirongo ya ADN nini, kandi gel isanzwe ya electrophoreis ntabwo ikwiye. Gisesengura kuri pulse gel electrophorei.Ni ibihe bibazo bindi wagize kuri agarose gel electrophorei? Tuzakora ubushakashatsi bwinshi kubayobora ejo hazaza.
Beijing Liuyi biotechnology Co., Ltd (Liuyi Biotech) nisosiyete yihariye yibanda ku bicuruzwa bifitanye isano na electrophoreis mu Bushinwa. Amateka yacyo atangira mu 1970 mugihe Ubushinwa bwari butarinjira mu ivugurura no gufungura igihe. Binyuze mu iterambere, Liuyi Bitotech ifite ikirango cyayo, kizwi ku izina rya Liuyi Brand ku isoko ry’imbere mu bicuruzwa bya electrophorei.
Ikirango cya Liuyi gifite amateka yimyaka irenga 50 mubushinwa kandi isosiyete irashobora gutanga ibicuruzwa bihamye kandi byujuje ubuziranenge kwisi yose. Binyuze mumyaka yiterambere, birakwiye guhitamo!
Utugingo ngengabuzima twa horizontal (tanks / chambers) ya Liuyi Biotech ni nziza cyane kandi igaragara neza. Hamwe nubunini butandukanye bwa gel tray, zirashobora kuzuza ibisabwa bitandukanye byubushakashatsi. Dufite itsinda ryacu rya tekiniki n'uruganda. Kuva mubishushanyo kugeza gukora, ibikoresho fatizo kugeza ibice byingenzi, dushobora kugenzura inzira zose. Urukurikirane rwa DYCP 31 ni urwa ADN electrophorei, nicyitegererezoDYCP-31BN, DYCP-31CN,DYCP-31DN, naDYCP-31E. Itandukaniro hagati yabo ni gel ingano nigiciro. Dutanga ingano yuzuye yibicuruzwa kubakiriya bacu. IcyitegererezoDYCP-32Cirashobora gukora gel nini 250mm * 250mm.
Hagati aho, turasaba amashanyarazi ya electrophoreisDYY-6C,DYY-6DnaDYY-10Ckuri selile ya electrophoreis (tanks / chambers) DYCP-31 na 32.
Niba ukeneye ibisobanuro byinshi kubyerekeye ibicuruzwa, nyamuneka sura uru rubuga kugirango ubone byinshi, kandi wakire neza kutwandikira ukoresheje imeri kugirango utumenyeshe icyo ushaka, urebe niba dushobora kuguha ibisubizo kuri wewe.
Kubindi bisobanuro kuri twe, nyamuneka twandikire ukoresheje imeri[imeri irinzwe], [imeri irinzwe].
Igihe cyo kohereza: Gicurasi-09-2022