Site Directed Mutagenesis Problem
Posted 03 March 2005 - 07:45 PM
Posted 20 February 2009 - 03:30 AM
Posted 21 February 2009 - 02:22 AM
Do you have only one colony? Try other colonies. You will get a right clone. I guess the additional 82bp insertion may result from your mutagenic primers. Both primers may not only perfectly match but also partially match each other. The partial match makes your insertion bigger.
have you sequenced the original vector to verify if this mutation is present. Also sequence other colonies, hopefully they are fine.