Chilling Bacterial Cultures Before Centrifugation
Posted 07 February 2009 - 12:03 PM
Posted 07 February 2009 - 06:36 PM
instead I believe the problem you are experiencing could be caused by
1 - too vigorous treatment (shaking) of the cell solution, once the cells are lysed with the lysis solution. This can shear the plasmid. The cell pellet has to be completely resuspended prior to addition of the lysis solution. And the two solutions mixed together gently.
2 - possible contamination of the reagents.
3 - using an expression strain rather than a cloning strain to prepare the plasmid.
Posted 07 February 2009 - 07:09 PM
Posted 08 February 2009 - 10:09 AM
So, what other steps are critical or have you noticed that if you do something wrong leads to a "dirty" prep?