The Easiness in Finding the N-Terminal

I was really shocked yesterday when I accidentally used the wrong primer pair to amplify the NT region. It was supposed to generate a 108 bp fragment out of the total NT region. Instead, I obtained almost 80% of the total region by using the primer pair to amplify the NT. Interestingly, the pE2-MaSpCL in the plasmid should not contain the full NT region or else I should have mistakenly used the wrong primer during the LD-PCR last time, thusmanaged to amplify the whole NT region as well. But nevertheless, if the whole NT presents in the insert plasmid, that would be just wonderful. And so far, I found it was very easy to amplify the NT region compared to that of CT. In fact, I managed to procure several bands including the GOI of extended NT, yet to be sequenced. Crossing fingers for positive result! Yosh~~