Alternatively, PCR can be performed using a Forward Primer that is designed to include the T7 promoter (with a few bases upstream) and the G to A change in the initiating nucleotide as well as some downstream sequence. Design the Reverse Primer to anneal to the region where your sequence will be terminated.
FAQ: How can I change the initiating sequence of my dsDNA template?
Alternatively, PCR can be performed using a Forward Primer that is designed to include the T7 promoter (with a few bases upstream) and the G to A change in the initiating nucleotide as well as some downstream sequence. Design the Reverse Primer to anneal to the region where your sequence will be terminated.