Figure 5

Cleavage of λ DNA (48,502 bp) under TaqII specificity changing conditions. (A) Complete TaqII cleavage pattern λ DNA under affinity star maximizing conditions (Methods). Samples of 1 μg λ DNA (=0.32 pmol recognition sites), digested under various conditions, were electrophoresed on 1% agarose/TBE gel. The TaqII/SIN/DMSO cleavage products were subsequently shotgun cloned to determine affinity star specificity (Methods). Lane M1, Fermentas 100 bp DNA Ladder (selected bands marked); lane M2, Fermentas 1 kb DNA Ladder (selected bands marked); Lane K, undigested λ DNA; lanes 1–4, λ DNA digested with TaqII: lane 1, without SIN and DMSO; lane 2, with SIN, no DMSO; lane 3, with 20% DMSO, no SIN; lane 4, with SIN and 20% DMSO. (B) Insert size distribution of 160 clones randomly selected from TaqII SIN/DMSO-generated λ DNA library in pUC19 vector (see Methods, Figures 2, 3 and 6). The average insert size of the library was estimated to be 160 bp.