Kwirinda polyacrylamide gel electrophorei

· polyacrylamidegel igomba na acrylamide monomer, polymerisation itangira ibikoresho, catalizator, nuburenganzira bwumunyu na buffer bivanze hamwe.

· acrylamidena BIS (N, N '- methylene double acrylamide) nuburyo bwa monomer form gel matrix.

· Amonium persulfate itangire ifata polymerisiyasi.Gukora kole bisaba 10% yumuti wa ammonium persulfate wateguwe mumazi.Amenshi mu makuru yerekanaga ko akeneye gukoreshwa neza.Nyamara, igisubizo cya 10% kirashobora gushyirwa kuri 4 ℃ ibyumweru byinshi nta gutakaza ibikorwa.Kora kugeza kuri ml 10 hanyuma ujugunye mugihe kole yananiwe guteranya.

Inama: Ijanisha ryaacrylamidemugukurikirana kole hamwe na protein glue ntabwo arimwe.Niba ukoresheje primaire acrylamide: BIS igisubizo, menya neza kubona icupa ryiza.

· TEMED (N, N, N ', N' - tetramethyl ethylenediamine) ni umusemburo, mumacupa yumukara, ashyirwa muri firigo.Ongeramo mbere yo gusuka kole.

· Polyacrylamide electrophoreis yakoreshejwe ikirahuri muri electrophoreis igomba gukaraba mbere na nyuma yigihe cyose.Nyuma ya electrophoreis, oza ukoresheje brush yoroshye hamwe nigitambara mumazi ashyushye yisabune, kwoza amazi yatoboye hanyuma uhagarare byumye.

· Ubushuhe hamwe n ivumbi birashobora gutera hamwe na polymer yuzuye.Mbere ya electrophoreis, kwoza isahani yikirahure hamwe nogusukura ibirahure hanyuma uhanagure hamwe na brush yoroheje.Karaba n'amazi yatoboye hanyuma wumishe neza ukoresheje impapuro.Kwoza hamwe na 70% Ethanol mbere yo guhanagura impapuro bifasha kweza no kwihuta byumye.Ongeraho ingero za acrylamide zikurikiranye: BIS, amazi, igisubizo cya buffer, ammonium persulfate, TEMED.Shyira neza hanyuma usuke ako kanya.

Ntabwo ari ngombwa gutesha polyacrylamide mbere ya polymerisation.(Acrylamide yahoze ishyirwa mu cyuho kugirango ikureho ibibyimba kuko ogisijeni ibuza polymerisiyasi.)

Gorizontal glue point sample sample.

· Shyira urupapuro rwumukara mumasanduku yo hepfo, inyuma yumukara kugirango ukore umwobo wintangarugero kugirango ubone neza.
· Gle tank yuzuye buffer, hejuru ya colloid.
· Niba inkombe ifite itara, fungura amatara, reka urumuri rumurikire.Shushanya icyitegererezo muri pipette.
· Gukoresha ibikoresho byikora byikora.
· Muri 10-200 mu 1 ingingo yimuka irashobora gukoreshwa mubice byinshi kurugero.Kubuto buto bw'icyitegererezo (munsi ya 10μ1), umutwe muremure wa pipette ukoreshwa mugukurikirana kole biroroshye.

· Gutobora kwibizwa gusa muri sample, guhumeka buhoro buhoro amazi.Icyitegererezo gishobora kugaragara gifatanye na glycerine, kandi kuvoma byihuse birashobora gukurura umwuka mwinshi mumutwe wa pipette.

· Icyitegererezo nyuma yo guhumeka umutwe wa pipetting, bizimura umutwe wamazi witonze kuruhande rwumuyoboro cyangwa guhanagura impapuro zonsa umutwe wamazi hanze yigitonyanga.Witondere kutanyunyuza icyitegererezo.

Shira icyitegererezo mu mwobo w'icyitegererezo

· Igikoresho cyo kuvoma kugirango ugumane umuvuduko muke, kora ingero zigenda buhoro hejuru yumutwe wamazi.
· Umutwe wumuyoboro winjijwe muri buffer, hejuru gato gato yumwobo, komeza umuvuduko mwiza.Isonga ya pipette irashobora kwinjizwa mumwobo muto.
· Buhoro buhoro kandi buhoro buhoro ibyitegererezo hanze.Umuyoboro wa pipette ushyizwe hejuru yicyitegererezo, kandi icyitegererezo kizarohama mu mwobo.Reka icyitegererezo kurohama kugirango wuzuze icyitegererezo aho gusunika.
· Iyo igitonyanga cyanyuma cyicyitegererezo gifite umutwe wamazi, amazi azimuka mukuguru kwa kabiri, azamura buhoro buhoro umuvuduko wimuka, wimuke uva muri buffer

Nigute wakora icyitegererezo cya kole?

· Uhagaritse kole point ya sample yibyobo byakozwe hagati yibice bibiri byikirahure.Muburyo bworoshye cyane, umutwe wa pipette ntushobora no kwinjizwa hagati yamasahani abiri.Reba glycerine!Shira umutwe wa pipette hejuru yumwobo wicyitegererezo hanyuma icyitegererezo kizarohama mu mwobo.
· Ingingo y'icyitegererezo mbere, menya neza ko ushira vertical point ya polypropilene acyl gel sample umwobo wogejwe neza.Koza acrylamide idashyizwe hamwe n'amazi ashobora kugaragara munsi yumwobo wintangarugero.Amazi arashobora gutuma umwobo wintangarugero uba muto cyane.Koresha inshinge ya 25ml cyangwa 50ml hamwe nurushinge rwa 18.Suka muri buffer ya electrophoreis hanyuma usukure neza umwobo w'icyitegererezo.
· Birashobora kugorana kubona umwobo wicyitegererezo, ariko kimwe, ibisigaye biroroshye.Niba hari umwobo urenze, sample ya buffer irashobora kugeragezwa hamwe n'ubururu bwa bromophenol.