Find content
Not Telling
Jerry, I'm not sure what the CutSmart buffer is made of, but "super buffers" can be very effective and have been around for a long time. An example is Mike McClelland's old KGB (Potassium Glutamate) buffer: http://www.ncbi.nlm....C334647/?page=1
#153015 What is NEB's CutSmart Buffer?
Posted
on 27 March 2013 - 06:11 AM
#147891 pEGFP C-1/N-1 Cloning
Posted
on 12 January 2013 - 05:05 AM
can you do a double digest to re-confirm your insert is there? I have worked with N2 and N3, and your protocol looks fine to me. One thing that I suspect is that your insert itself could be toxic? or your competent cells are not viable enough due to a possible break down of your freezer. Trust me this has happened to us like more than 5-6 times that during weekend we had no electricity and nobody knew about it. These cells might grow in broth again, but are not suitable for transformation anymore. Kan plates also last for long time, unlike amp plates that can't be used after a month.
