Trouble with Phosphorylation/Ligation of a blunt-ended CRISPR plasmid. Any troubleshooting ideas?

I have been trying to phosphorylate and ligate a blunt-ended linear plasmid after PCR. It remains linear. Any suggestions for troubleshooting? I have tried 10 min at 70 degrees followed by ice for...

16 March 2017 3,727 3 View