Are You There NT & CT Termini?

After confirming the presence of empty plasmid in the cell, I further evaluated the presence of the MaSpCL insert just in case prior to long fragment sequencing. I did not want the previous situation occurred again whereby the sequencing of a 11 kb amplicon; believed to be the right MaSpCL resulted in an unknown sequence thus, wasting money and time. So, I designed two sets of primer targeting the NT and CT termini of the hypothetical MaSpCL and run the PCR. As expected, the NT terminus was obtained together with the CT. I was so relieved because now I am confident that the insert is actually the real putative MaSpCl. Now, I am preparing the insert plasmid in large amount for sequencing! Crossing fingers to this!

                                       The designed primers yielded that NT terminus of interest.