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== 阶段二 制胶 == *'''组装制胶装置''' *'''配置分离胶(说明书)''' *'''灌胶:用1 ml移液枪加下层胶,沿一边灌至绿色下缘线,再加左右滑动上层胶至玻璃板边缘,立即慢慢插入梳子,上下层 胶分界线与梳子底部至少大于0.5 cm,放置37 ℃烘箱20 min,等待胶凝固同时配置电泳缓冲溶液''' *'''转移至电泳槽中,短板向内,加入电泳缓冲液至2 gel刻度线,取出玻璃板放入电泳缓冲液中,补满小槽内缓冲液,加大槽缓冲液4 gel处,两槽需有液面高度差,再慢慢拔出梳子''' *'''上样:两边孔弃用或者加Loading buffer,避免边缘效应,上样前可用电泳缓冲液稀释各样品至体积一致,Marker和Loading buffer以1:1混合,Marker 8 ul + 缓冲液 2 ul,不上样的孔加Loading buffer 10 ul''' *'''电泳:点 Manual 点 V,90 V 恒压,小槽内有许多气泡向上跑,RUN,进入分离胶越50 min,进入分离胶后可调至 150 V,等待时配置转膜缓冲液'''
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