I phosphorylated the primers and followed by PCR. I did the blunt end self ligation using T4 DNA ligase. After ligation I got colonies. I purified the plasmid and sent them for sequencing. The results shows that there is a truncate at the ligation point. It did not end to end ligate while it ligate with 3 nucleotide truncated.
correct one should be:
But I got like this:
there are three nucleotide truncated.
Any suggestions why it can happened?
Edited by Dee, 16 July 2010 - 03:49 AM.