Hi,can anybody suggest the method to check the orientation of my gene cloned into pPic9 vector , my gene size is 1.5kb and it is cloned at Eco r1 site.
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1 reply to this topic
Posted 14 August 2002 - 06:42 AM
To check your orientation try and find a combination of restriction enzyme sites, one in your vector, and one in your insert near to one end. You can then work out which way the insert has gone in by by doing a double digest, one pattern of bands will tell you the correct orientation, the other will tell you if its in the wrong way.