Ingamba za polyacrylamide Gel electrophores

· PolyacrylalideGel agomba kubuza bwa Acrylamude, polymeration polymerimiation gutangira ibikoresho, umusemburo, nuburenganzira bwumunyu na buffer bivanze hamwe.

· acylamidena bis (n, n '- methylene acrylamide ebyiri) ni monomer ikora gel matrix.

· Ammonium yerekanaga itangira inzira yo gufatira. Gutegura kole bisaba 10% ya memonium yerekana amakuru yateguwe mumazi. Amakuru menshi yerekanaga ko ari ngombwa gukoresha neza. Ariko, igisubizo cya 10% kirashobora gushyirwa saa 4 ℃ mu byumweru byinshi nta gutakaza ibikorwa bikomeye. Kora kugeza kuri ml 10 hanyuma ujugunye iyo kole inanirwa guterana.

Inama: ijanisha ryaacylamideMu gukurikiranya kole na koroue ya poroteyine ntabwo arimwe. Niba ukoresheje acrylamide ya firime: Bis Igisubizo, menya neza kugirango ubone icupa rikwiye.

· Temed (n, n, n, n ', tetramethyl etylesesinine) ni umusemburo, mu icupa ryijimye, rishyirwa muri firigo. Ongeraho gusa mbere yo gusuka kole.

· Polyacrylandide ya electrophos yakoresheje ikirahuri muri electrophoresis igomba gukaraba mbere na nyuma yigihe gito. Nyuma ya electrophoresis, gukaraba hamwe no gukaraba byoroshye n'amazi ashyushye, kwoza amazi yatoboye kandi uhagarare kumema.

· Ubushyuhe n'umukungugu birashobora gutera hamwe na polymer yuzuye. Mbere ya electrophoressis, humura isahani yikirahure hamwe nibirahure hanyuma uhanagure hamwe na brush yoroshye. Gukaraba hamwe namazi yatoboye no gukama neza hamwe nimpapuro. Kwoza hamwe na 70% Ethanol mbere yo guhanagura impapuro zifasha gusukura no kwihuta. Ongeramo ingero za acylamide ikurikira: Bis, amazi, igisubizo buffer, amonimonipe ya amonium, birimo. Kunyeganyeza neza hanyuma uhite usuka.

Ntabwo ari ngombwa kuri delyacrylamide mbere ya polymeiriation. .

Horizontal glue ingingo yibitekerezo.

· Shyira urupapuro rwirabura mumasanduku yo hepfo, umukara inyuma kugirango ukore umwobo wintangarugero kugirango ubone neza.
· Glue Tank yuzuye buffer, hejuru ya colloid.
· Niba inkombe ifite urumuri, fungura amatara, reka urumuri rumurikire. Shushanya icyitegererezo muri pipette.
· Ikoresha igikoresho cya pipa.
· Muri 10-200 mu 1 Ingingo ya Lifes irashobora gukoreshwa mubiganiro byinshi kurugero. Kubintu bito cyane byingoro (munsi ya 10μ1), umutwe muremure wa pipette ukoreshwa mugukurikirana kole nibyinshi byoroshye.

· Pipetting gusa yibizwa murugero, guhumeka buhoro buhoro amazi. Icyitegererezo kirashobora kugaragara ko na Glycerin, kandi kuvoma byihuse birashobora gushushanya umwuka mu mutwe wa pipette.

· Icyitegererezo nyuma yo guhumeka umutwe, bizamura umutwe wamazi witonze kuruhande rwumuyoboro cyangwa guhanagura impapuro zonsa umutwe hanze yimbwa. Witondere kutanywa icyitegererezo.

Shira icyitegererezo muruzitiro rwicyitegererezo

Igikoresho cyo gushushanya kugirango ukomeze igitutu gito, kora ingero zigenda zigenda zigenda zirenga umutwe.
· Umutwe wa pipetting winjijwe muri buffer, hejuru gato kurenza umwobo, kubungabunga igitutu cyiza. Isonga rya pipette irashobora kwinjizwa mu mwobo muto.
Buhoro kandi buhoro kandi buhoro buhoro ingero ziguruka. Impanuro ya pipette yashyizwe hejuru yikigo cyicyitegererezo, kandi icyitegererezo kizarohama mu mwobo. Reka icyitegererezo kirohama rwuzuze umwobo wicyitegererezo aho gusunika.
· Igihe cyanyuma cyicyitegererezo hamwe numutwe wamazi, amazi azimuka kumuguru wa kabiri, uzamure buhoro buhoro, wimuke muri buffer

Nigute ushobora gukora ingendo zihagaritse?

· Vertical glue ingingo yibishyimbo byimpyiko zakozwe hagati yikirahure. Muri kole nto cyane, umutwe wa pipette ntushobora no kwinjizwa hagati yamasahani abiri yikirahure. Reba Glycerin! Shira umutwe wa pipette hejuru yicyitegererezo kandi icyitegererezo kizarohama mu mwobo.
· Tekereza icyitegererezo mbere, menya neza gushira ingingo ihagaritse ya polypropylene acyl gel icyitegererezo cyuruziga rwuzuye. Kwoza acylamide ya acrymedide hamwe namazi ashobora kugaragara munsi yumwobo wicyitegererezo. Amazi arashobora gukora umwobo w'icyitegererezo cyane. Koresha syringe ya 25ml cyangwa 50ml hamwe numushinge 18. Suka muri electrophoresis buffer kandi witonze witonze umwobo wicyitegererezo.
· Birashoboka ko bigoye kubona umwobo wicyitegererezo, ariko kuri kimwe, ibisigaye biroroshye. Niba hari umwobo urenze, icyitegererezo gishobora kugeragezwa hamwe na bromophenol ubururu.