EcoRI cloning giving only inverted inserts!!??
Posted 30 September 2010 - 02:12 PM
To verify the orientation of the insert I´m using an enzyme that cuts one time in the vector and one time in the insert. The difference is very clear (400pb vs 1200pb).
Someone has a clue? Or am I very unlucky?
Posted 30 September 2010 - 02:21 PM
Posted 30 September 2010 - 02:30 PM
The vector is a lentiviral vector with a SFFV promoter, generally used for hematopoietic cells. Is this promoter active in E. coli??? Maybe a "residual" activity could cause this problem?
Thanks for the answer!
Posted 30 September 2010 - 03:16 PM