Kusamala kwa polyacrylamide gel ectophmoresis

· polyacrylamideGel Ayenera kutomer, polymerization kuyambiranso zinthu, chothandizira, ndi ufulu wa mchere ndi osakaniza.

· acylamideNdipo Bis (n, n '- methylene flue for acamide) ndi fomu ya monomer eltrix.

Ambumoni pemphanimium kuyamba kutsatira njira zolaula. Kupanga kwa guluu kumafuna 10% ammonium pemphangote yankho lomwe lakonzedwa m'madzi. Zambiri mwazidziwitso zomwe zimawonetsa kufunikira kogwiritsa ntchito. Komabe, yankho la 10% limatha kuyikidwa pa 4 ℃ kwa milungu ingapo popanda kutaya ntchito. Pangani mpaka 10 ml ndikutaya pomwe guluu limalephera kuphatikizika.

Nsonga: kuchuluka kwaacylamideMu chosindikizira guluu ndi mapuloteni mapuloteni sichofanana. Ngati mukugwiritsa ntchito ma acrylamide premede: Bis Solution, onetsetsani kuti mwapeza botolo lamanzere.

Mosachita manyazi (n, n, n ', n' - TETRAMTHYL Ethyleniamine) ndi chothandizira, m'botolo yofiirira, yoyikidwa mufiriji. Onjezani musanalowe gululu.

Ma elel`nacryylamide ectropeteces ogwiritsa ntchito ma electrophoresis ayenera kutsukidwa kale komanso pambuyo pake. Pambuyo pa electrophoresis, sambani ndi burashi yofewa ndi nsalu yamadzi otentha a sopo, nadzatsuka ndi madzi osungunuka ndikuyima kuti awume.

Ndipo chinyontho ndi fumbi zingayambitse polocha. Asanayambe electrophoresis, yeretsani mbale yagalasi yokhala ndi magulasi ndikupukuta ndi burashi yofewa. Sambani ndi madzi osungunuka ndikuwuma bwino ndi pepala. Kukulitsa ndi 70% ethanol musanapumire ndi pepala kumathandizira kuyeretsa ndi kufulumira kuyanika. Onjezani zitsanzo za ma acrylamide motere: Bis, madzi, yankho la buffer, ammonium Perulpatete, amamwa. Gwedezani bwino ndikutsanulira nthawi yomweyo.

Sikofunikira ku Degrass Polyacrkala lisanafike polymerization. (Acrylamide adayikidwa mu vacuum kuti muchotse thovu chifukwa oxygen amaletsa polymerization.)

Nsonga zopingasa zikuluzikulu.

Chida cha pansi, chakuda kuti chikhale chodulira kuti muwone bwino.
Tankinkinki yodzaza Buffer, wokha wa Colloid.
Ngati m'mphepete ili ndi nyali, iyatse nyali, kuwala kunyezimira. Jambulani nyemba mu pipette.
Chida chojambulira zokha.
4-200 mu 1 Madzi osunthira akhoza kugwiritsidwa ntchito m'malo ambiri mwa zitsanzo. Kwa mabowo ang'onoang'ono kwambiri (ochepera 10μ1), mutu wautali wa piya womwe umagwiritsidwa ntchito kukhomera ndikosavuta.

Kupukutira kumangomizidwa mu zitsanzo, kumapangitsa pang'ono pang'onopang'ono. Chitsanzo chimatha kuwoneka ngati chomata ndi glycerin, ndipo mpomwa msanga amatha kujambula thovu mu mutu wa ma pipette.

Zitsanzo zikakhala zikhulupiliro zitatha, zisunthiratu pang'ono pamphepete mwa chitoliro kapena kupukusa pepala lamadzimadzi kunja kwa madontho. Samalani kuti musayankhe zitsanzo.

Ikani zitsanzozo mu dzenje

Chida chofananira kuti musunge pang'ono, pangani zitsanzo pang'ono pang'onopang'ono kusuntha mutu.
Mutu wa pakhosi womwe umayikidwa mu buffer, wokwezeka kwambiri kuposa mabowo owoneka, samalani. Nsonga ya pipette ikhoza kuyikidwa mu dzenje laling'ono.
Mosadukiza ndi kusakhazikika. Umu wa pipette umayikidwa pamwamba pamtunda dzenje, ndipo zitsanzozo zimira mdzenje. Lolani zikondweretse kuti mudzaze mabowo m'malo mokankhiramo.
· Khumi womaliza ndi mutu womaliza ndi mutu wamadzimadzi, madziwo amapita ku mwendo wachiwiri, pang'onopang'ono kwezani madzi kusuntha, tulukani kuchokera ku buffer

Kodi mungachite bwanji magawatala ofukula?

Miyambo yolunjika mabowo amapangidwa pakati pa zidutswa ziwiri zagalasi. Mu guluu wowonda kwambiri, mutu wa piya umatha kuyikanso pakati pa mbale ziwiri zagalasi. Yang'anani glycerin! Ikani mutu wa pipette pamwamba pa dzenje ndi zitsanzozo zimira mdzenje.
Samizani zitsanzo zisanachitike, onetsetsani kuti mwayika zolimba za acyl gels gel osakaniza ndi zoyera. Sambani ma acrylamized acpomide oyimitsa ndi madzi omwe angawonekere pansi pa dzenje. Madziwo amatha kupangitsa kuti dzenje lizikhala laling'ono kwambiri. Gwiritsani ntchito syringe kapena 50ml kapena suri ya 18-gauge. Kutsanulira mu bufferophorsus buffer ndikupukutira mosamala.
Ingakhale yovuta kuwona dzenje, koma chimodzimodzi, enawo ndi osavuta. Ngati pali mabowo owonjezera, mawonekedwe osungidwa amatha kuyesedwa ndi blomapeni buluu.