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010咦

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[求助] SDS-PAGE

用BL21表达目的蛋白后,取未加IPTG诱导的菌液1ml,离心倒掉上清,将80ul的1×PBS和10ul的PMSF混匀后加入沉淀中,将沉淀重悬以后加入20ul的loading buffer沸水浴10分钟,然后离心10分钟,跑SDS-PAGE点样时,有的样品不会下沉到胶孔中,有的样品却可以,是哪个处理步骤出现问题呢?怎么解决?已经排除不是胶的问题。

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Glenice Y

新虫 (正式写手)

应该是某些菌体浓度过高,导致菌体粘稠不好上样,你可以多加一些PBS按照你的方案在做,相当于扩大体积

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