我的lab 买了一株还特定clone 之菌 该 vector 为 pSin-EF2-Pur 遽我所知此载体将用来做感染细胞做病毒用 买了的菌株是 STBL3 查了一下这株菌 含有endonuclease 1 (endA1+) : Reference: 摘录自invitrogen 的 Kit: pLenti6/V5 Directional TOPOR Cloning Kit http://tools.invitrogen.com/content/sfs/manuals/plenti6v5dtopo_man.pdf Stbl3? E. coli is wild type for endonuclease 1 (endA1+). When performing plasmid DNA isolation with commercially available kits, ensure that Solution I of the Lysis buffer (often called Resuspension Buffer) contains 10 mM EDTA to inactivate the endonuclease to avoid DNA nicking and vector degradation. 当我养菌, 并用 geneid 的 mini kit 抽载体时, 载体若泡在限制梅的Buffer就会被切烂 後来我听说只要把载体转染至 DH5alpha 就不会有这样的现象 Reference: Problems with STBL3 cells - Digesting DNA from STBL3 cells http://www.protocol-online.org/biology-forums/posts/11640.html 想请问过来人的经验: 1. STBL3 这株菌有特殊用途吗? 为甚麽要用这株菌? 2. 我要操作vector pSin-EF2-Puro 以便将来制作病毒感染细胞用 可以转染到DH5alpha 来操作就好吗? 感谢~~ --

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