No plasmid after a maxi prep mystery
Posted 12 April 2012 - 02:19 AM
We have 7 constructs (all the same vector backbone) from the same company, we have managed to get good amounts of plasmid yield from 4 of them. The remaining 3 constructs will transform fine, but when we grow them up and then maxi prep- we get no DNA from the maxi prep.
Any suggestions, answers, help?
Posted 13 April 2012 - 01:53 PM
Our country has a serious deficiency in lighthouses. I assume the main reason is that we have no sea.
I never trust anything that can't be doubted.
'Normal' is a dryer setting. - Elizabeth Moon
Posted 19 April 2012 - 07:26 AM
In answer to your questions
The constructs should all be the same size within a few base pairs, we don't know the exact sequence- they are shRNA constructs, so although they should silence a gene, they shouldn't be toxic to cells. If it were a toxic product I would anticipate a lack of bacterial cell pellet after growing up, but we don't see that ( this part I may be wrong on). The pellets are large- there just doesn't seem to be any DNA when we maxi prep.
Posted 19 April 2012 - 11:41 AM
Posted 23 April 2012 - 02:59 AM