FAQ: Is it necessary to add a “G” into the oligo following the T7 promoter sequence?
At least one G, directly downstream of the “TATA” of the T7 promoter
sequence is required for efficient transcription. This G will be the
+1 of the transcript. If the 20 nucleotide target-specific sequence
selected has a G at the 5’ end of the 20 nucleotides then it is not
necessary to add an additional G as the G within the target-specific
sequence will serve as the +1.