Polyacrylamide Gel Electrophoresis
Gel electrophoresis wata hanya ce ta asali a cikin dakunan gwaje-gwaje a duk fannonin nazarin halittu, suna ba da izinin rabuwar macromolecules kamar DNA, RNA da sunadarai.Kafofin watsa labarai na rarrabuwa daban-daban da hanyoyin suna ba da damar ɓangarorin waɗannan ƙwayoyin cuta don rabuwa da kyau ta hanyar amfani da halayensu na zahiri.Don sunadaran musamman, polyacrylamide gel electrophoresis (PAGE) galibi shine dabarar zaɓi.
PAGE wata dabara ce da ke raba macromolecules kamar sunadaran gina jiki bisa la’akari da motsin su na electrophoretic, wato ikon masu nazari don matsawa zuwa na’urar lantarki sabanin caji.A cikin PAGE, ana ƙayyade wannan ta caji, girman (nauyin kwayoyin halitta) da siffar kwayar halitta.Analytes suna motsawa ta hanyar pores da aka kafa a cikin gel polyacrylamide.Ba kamar DNA da RNA ba, sunadaran suna bambanta iko bisa ga haɗin amino acid, wanda zai iya yin tasiri akan yadda suke gudana.Amino acid kirtani na iya samar da sifofi na biyu waɗanda ke tasiri ga girmansu da kuma yadda suke iya motsawa ta cikin pores.Don haka yana iya zama abin sha'awa a wasu lokuta don cire sunadaran sunadaran kafin electrophoresis don daidaita su idan ana buƙatar ƙarin ƙimar girman daidai.
SDS PAGE
Sodium-dodecyl sulfate polyacrylamide gel electrophoresis wata dabara ce da ake amfani da ita don raba kwayoyin furotin na talakawa 5 zuwa 250 kDa.Sunadaran sun rabu ne kawai akan nauyin kwayoyin su.Sodium dodecyl sulfate, anonionic surfactant, ana ƙara shi a cikin shirye-shiryen gels wanda ke rufe cajin abubuwan da ke cikin samfuran furotin kuma yana ba su irin wannan cajin zuwa rabon taro.A cikin kalmomi masu sauƙi, yana hana sunadarai kuma yana ba su caji mara kyau.
PAGE na asali
Native PAGE wata dabara ce da ke amfani da gels marasa ƙima don rarrabuwar sunadaran.Ba kamar SDS PAGE ba, babu wani wakili da aka ƙara a cikin shirye-shiryen gels.A sakamakon haka, rarrabuwar sunadaran suna faruwa ne bisa ga caji da girman sunadaran.A cikin wannan fasaha, juzu'i, naɗewa da sarƙoƙin amino acid na sunadaran sune abubuwan da rabuwar ke dogara da su.Sunadaran ba su lalace ba a cikin wannan tsari, kuma ana iya dawo dasu bayan kammala rabuwa.
Ta yaya polyacrylamide gel electrophoresis (PAGE) ke aiki?
Babban ka'idar PAGE ita ce raba masu nazari ta hanyar wucewa ta cikin pores na gel polyacrylamide ta amfani da wutar lantarki.Don cimma wannan, an haɗa haɗin acrylamide-bisacrylamide polymerized (polyacrylamide) ta ƙari na ammonium persulfate (APS).Halin, wanda tetramethylethylenediamine (TEMED) ke haɓakawa, yana samar da tsari kamar tsari tare da pores ta hanyar da masu bincike zasu iya motsawa (Hoto 2).Mafi girma yawan adadin acrylamide da aka haɗa a cikin gel, ƙananan girman pore, saboda haka ƙananan sunadaran da za su iya wucewa.Rabon acrylamide zuwa bisacrylamide shima zai yi tasiri ga girman pore amma ana kiyaye wannan sau da yawa.Ƙananan ƙananan ƙananan ƙwayoyin cuta kuma suna rage saurin da ƙananan sunadaran suna iya motsawa ta hanyar gel, inganta ƙudurin su da kuma hana su gudu zuwa cikin buffer da sauri lokacin da ake amfani da halin yanzu.
Kayan aiki don Polyacrylamide Gel Electrophoresis
Gel Electrophoresis Cell (Tank/ Chamber)
Tankin gel na polyacrylamide gel electrophoresis (PAGE) ya bambanta da tankin gel na agarose.Tankin gel na agarose yana kwance, yayin da tankin PAGE yana tsaye.Ta hanyar cell electrophoresis a tsaye (tanki / ɗakin), ana zuba gel na bakin ciki (yawanci 1.0mm ko 1.5mm) a tsakanin faranti biyu na gilashi kuma a saka shi ta yadda kasan gel ɗin yana nutsewa a cikin ɗaki ɗaya kuma saman yana nutsewa a cikin buffer. a wani dakin.Lokacin da ake amfani da halin yanzu, ƙaramin adadin buffer yana ƙaura ta gel daga ɗakin sama zuwa ɗakin ƙasa.Tare da ƙwanƙwasa masu ƙarfi don ba da tabbacin taron ya tsaya a madaidaiciyar matsayi, kayan aikin yana sauƙaƙe tafiyar gel mai sauri tare da sanyaya ko da yana haifar da ƙungiyoyi daban-daban.
Beijing Liuyi Biotechnology Co., Ltd. (Liuyi Biotechnology) yana kera nau'ikan nau'ikan nau'ikan ƙwayoyin polyacrylamide gel electrophoresis (tankuna / ɗakuna).Samfuran DYCZ-20C da DYCZ-20G sel electrophoresis ne a tsaye (tankuna/ɗakuna) don nazarin jerin DNA.Wasu daga cikin sel electrophoresis na tsaye (tankuna / ɗakunan) sun dace da tsarin toshewa, kamar samfurin DYCZ-24DN, DYCZ-25D da DYCZ-25E sun dace da tsarin tsarin Blotting Western DYCZ-40D, DYCZ-40G da DYCZ-40F. waɗanda ake amfani da su don canja wurin kwayoyin furotin daga gel zuwa membrane.Bayan SDS-PAGE electrophoresis, Western Blotting wata dabara ce don gano takamaiman furotin a cikin cakuda furotin.Kuna iya zaɓar waɗannan tsarin gogewa bisa ga buƙatun gwaji.
Electrophoresis Power Supply
Don samar da wutar lantarki don tafiyar da gel, kuna buƙatar wutar lantarki ta electrophoresis.A Liuyi Biotechnology muna samar da kewayon samar da wutar lantarki ga duk aikace-aikace.Model DYY-12 da DYY-12C tare da babban barga irin ƙarfin lantarki da na yanzu iya saduwa da high ƙarfin lantarki da ake bukata electrophoresis.Yana da aikin tsayawa, lokaci, VH da aikace-aikacen mataki-mataki.Sun dace don aikace-aikacen IEF da DNA sequencing electrophoresis.Don amfanin furotin na gabaɗaya da aikace-aikacen electrophoresis na DNA, muna da samfurin DYY-2C, DYY-6C, DYY-10, da sauransu, waɗanda kuma ke samar da wutar lantarki mai zafi tare da ƙwayoyin electrophoresis (tankuna / ɗakuna).Ana iya amfani da waɗannan don aikace-aikacen electrophoresis na tsakiya da ƙananan ƙarfin lantarki, kamar don amfani da ɗakin karatu na makaranta, dakin gwaje-gwaje na asibiti da sauransu.Ƙarin samfura don samar da wutar lantarki, da fatan za a ziyarci gidan yanar gizon mu.
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Nassoshi don Menene polyacrylamide gel electrophoresis?
1. Karen Steward PhD Polyacrylamide gel electrophoresis, Yadda yake Aiki, Bambancin Fasaha, da aikace-aikacen sa
Lokacin aikawa: Mayu-23-2022