WangLab:Deletion or Insertion of Amino Acids w/o the Involvement of RE sites

Deletion or Insertion of amino acids without the involvement of restriction enzyme sites

 * Design primers as indicated:




 * Run PCR as following (strategene mutagenesis kit):


 * Digest parental DNA with 0.5 ul DPNi for >2hr 37oC.
 * Gel purify the PCR product, do blunt ligation:


 * Do transformation