· polyacrylamidegel rinofanira neacrylamide monomer, polymerization yekutanga zvinhu, catalyst, uye kodzero yemunyu uye buffer musanganiswa pamwe chete.
· acrylamideuye BIS (N, N '- methylene double acrylamide) ndiyo monomer form gel matrix.
· ammonium persulfate tanga adhesive polymerization process.Kuumbwa kweglue kunoda 10% ammonium persulfate solution yakagadzirwa mumvura.Ruzhinji rweruzivo rwairatidza kudiwa kwekushandisa nesimba.Nekudaro, iyo 10% mhinduro inogona kuiswa pa4 ℃ kwemavhiki akati wandei pasina kurasikirwa kukuru kwekuita.Ita kusvika ku 10 ml uye urase kana glue yakundikana kuunganidza.
Zano: Chikamu cheacrylamidemukutevedzana glue uye protein glue haina kufanana.Kana uchishandisa premade acrylamide: BIS mhinduro, ita shuwa kuti wawana bhodhoro chairo.
· TEMED (N, N, N ', N '- tetramethyl ethylenediamine) inogadzirisa, mubhodhoro rebrown, yakaiswa mufiriji.Wedzerai musati madira glue.
· polyacrylamide electrophoresis yakashandiswa girazi mu electrophoresis inofanira kugezwa nguva isati yasvika uye yapera.Mushure me electrophoresis, geza nebhurashi rakapfava uye jira mumvura inopisa ine sipo, simbura nemvura yakasvibiswa uye umire kuti uome.
· unyoro neguruva zvinogona kukonzera ne hollow polymer.Pamberi pe electrophoresis, chenesa ndiro yegirazi negirazi yekuchenesa uye uipute nebhurashi rakapfava.Shamba nemvura yakasvibiswa uye uomese zvakakwana nepepa rekupukuta.Kusuka ne70% ethanol usati wapukuta nebepa kunobatsira kuchenesa nekukasira kuomesa.Wedzera sampuli dze acrylamide zvakatevedzana: BIS, mvura, buffer solution, ammonium persulfate, TEMED.Kuzunungusa zvakanaka wodira pakarepo.
Hazvina kudikanwa kuti degass polyacrylamide pamberi polymerization.(Acrylamide yaimboiswa muvacuum kuti ibvise mabubbles nekuti oxygen inotadzisa polymerization.)
Horizontal glue point sample tips.
· isa bepa dema mubhokisi rezasi, dema rekumashure kugadzira imwe yemuenzaniso gomba kuti unyatsoona.
· glue tangi rakazara buffer, pamusoro pecolloid.
· kana mupendero une chiedza, vhenekera mwenje, chiedza ngachivheneke colloid.Dhirowa muenzaniso mu pipette.
· kushandisa otomatiki pipetting mudziyo.
· mu10-200 mu 1 mvura inofambisa poindi inogona kushandiswa mune mazhinji emapoinzi pamuenzaniso.Kune maburi madiki emuenzaniso (asingasviki 10μ1), musoro wakareba wepipeti unoshandiswa kuteedzera glue uri nyore.
· kupinza pombi kungonyudzwa mumucherechedzo, kufema mvura inofamba zvishoma nezvishoma.Sample inogona kuita seyakanamatira neglycerin, uye kukurumidza kupomba kunogona kukwevera mabhuru emhepo mumusoro wepipette.
sampuli mushure mekufema musoro wepombi, inofambisa musoro wemvura zvinyoro-nyoro pamupendero wepombi kana kupukuta bepa rinoyamwa musoro wemvura kunze kwemadonhwe.Chenjerera kuti usayamwa sampuli.
Isa muenzaniso mugomba remuenzaniso
· pipetting mudziyo kuchengeta zvishoma kumanikidza, ita kuti sampuli zvishoma kufamba mafashama mvura musoro.
· Musoro wepombi wakapinzwa mubuffer, wakakwira zvishoma pane maburi enzvimbo, chengetedza kudzvanywa kwakanaka.Musoro wepipette unogona kuiswa mugomba duku.
· zvishoma nezvishoma uye zvishoma nezvishoma masampuli kunze.Iyo pipette tip inoiswa pamusoro peiyo poindi sampuli gomba, uye sampu inonyura mugomba.Rega samuenzaniso inyure kuzadza gomba remuenzaniso pane kusundira mukati.
· kana kamwe chete donho rekupedzisira remuenzaniso rine musoro wemvura, mvura inoenda kugumbo rechipiri, ichisimudza zvishoma nezvishoma kufamba kwemvura, kubuda kunze kwebuffer.
Maitiro ekuita vertical glue sampling?
· vertical glue point makomba maburi akaumbwa pakati pezvidimbu zviviri zvegirazi.Mune glue yakaonda kwazvo, musoro wepipette haugone kuiswa pakati pemahwendefa maviri egirazi.Tarisa glycerin!Isai musoro wepipette pamusoro pegomba remuenzaniso uye sampuli inonyura mugomba.
· poindi sampuro kare, iva nechokwadi chekuisa poindi yakatwasuka yepolypropylene acyl gel sampuli gomba rinogezwa rakachena.Geza iyo unpolymerized acrylamide uye mvura inogona kuoneka pazasi pegomba remuenzaniso.Mvura inogona kuita kuti gomba remuenzaniso rive diki zvakanyanya.Shandisa 25ml kana 50ml sirinji uye 18-gauge tsono.Dururira mu electrophoresis buffer uye nyatso penya gomba remuenzaniso.
· zvinogona kunetsa kuona gomba remuenzaniso, asi panguva imwe chete, mamwe ari nyore.Kana paine maburi akawandisa, iyo sampuli buffer inogona kuyedzwa nebromophenol yebhuruu.
Nguva yekutumira: Mar-31-2023