Figure 3.

Purity analysis of the fully deblocked 40b strand before and after treatment with RecJ and SSBP. (A) Two superimposed RP HPLC chromatograms of the 40b strand (200 pmol total) before (I) and after (II) purification with 60 units of RecJ and 0.5 μg of SSBP. Both samples were treated with 0.1 M NaOAc (pH 3.5) postreaction to demonstrate the purity of final detritylated product. (B) PAGE image of the same two samples before (I) and after (II) purification with the disappearance of truncated failure product. Gel analysis was performed on 15% PAGE with urea (120 V for 60 min). Both chromatograms were offset for the sake of clarity and have been moved along the x-axis; the control target FLP peak matched up with the target FLP peak in Figure 2. The larger peak for the untreated FLP-off sample is due to the addition of prematurely detritylated FLP [127.5 pmol + 35.4 pmol (Figure 2B)].