[求救] 有關gel extraction
看板Biotech (生命科學)作者blueashin (tRoMboNe pLaYer)時間15年前 (2010/03/29 14:38)推噓4(4推 0噓 24→)留言28則, 7人參與討論串3/3 (看更多)
想請教板上的各位先進
最近我有一條基因想要送讀序
可是一直無法以gel extraction的方式成功回收
但是因為pcr的產物 量實在是無法增加
所以只能在gel extraction方面加強
以下是我gel extraction的步驟:我使用的是V牌國產的回收kit
切下膠塊以500ul GEX buffer回溶
60 度C加熱10分鐘 反轉2-3次
通入column用vocume method
WN buffer500ul
WS buffer 500ul
13000rpm 25度c離心3分鐘
加入預熱60度C之EB或者 ddH2O(pH=7)
等待10分鐘
13000rpm 25度C離心3分鐘
以上步驟 不知道到底是哪理出問題了QQ還是說是他們的column有問題?
我測OD值都很低 有時候還測不到東西
縱使有測到 A260/230的值也很低
要做ligation實在有困難QQ
以上 懇請各位指教~
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