[求救] NHEJ assay 原理看不太懂
大家好
小弟最近在讀一篇paper 上面有做了NHEJ assay
大概就是說某個基因會加速NHEJ 把那個基因抑制之後發現細胞colony變少
做法是使用兩組細胞 一組control 一組siXXXX (那個基因)
然後另外使用G418去篩選 最後觀察到有G418 也有siXXXX的細胞 colony最少
原文是這樣說 :
Cells were transfected with a linearized vector
harboring a neomycin-resistant gene and an siRNA targeting XXXX (基因)
In the presence of neomycin, only cells that can
integrate the plasmid via NHEJ survive
所以我又去查了他transfect什麼plasmid
結果是這個 : BamHI-XhoI linearized pEGFP-C1
我不懂的是 這個plasmid看起來就是表現GFP 那他要怎麼篩出
只經由NHEJ存活下來的細胞?
不知道有沒有做過這個實驗的學長姊
我看好久還是不太懂
麻煩各位解惑了!!
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