小中大我是这样做的,取出海马组织,用0.25%胰酶37度消化20分钟,然后用吸管吹打,
100目的筛板过滤,取滤液1000转离心5分钟,去上清液加PBS吹打成悬液,但每次都不成功,不是细胞成团就是细胞碎了,请问是什么原因
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It seem that you have treated the tisssue with trypsin too long. Over trypsinization will lead cell death and cell DNA release. The released DNA will make your digestion or cells clump. To solve the problem, you can 1) shorten the enzyme digestion time; or 2) add DNase at the last five minutes of enzyme digestion to degrade the released DNA. Let me know if you have further problem.