Extra T7 Priming Site Extra T7 priming site Client presented a template for sequencing. Reaction with M13 reverse primer worked well.
Baseline is flat indicating template is clean.
Reaction with T7 primer had many N's and superimposed peaks, not usable sequence. Person processing the data thought there had been leakage from an adjoining lane on the gel. Remaining portion of sample was loaded on another gel and produced the same peak pattern. The template was tried with the M13 forward primer and produced good sequence.