Figure 5.

DmGen and HsGEN1 cut 5΄ flaps faster than HJs in a variety of reaction conditions. (A) Schematic showing the substrates and reaction buffers tested. The substrates shown in red and dark blue are those used in our experiments (‘Bellendir oligos’), and those substrates in magenta and cyan are from the previous published study on HsGEN1 (‘Rass oligos’) (9). The sequences of central nucleotides near the branch points are shown in black. The buffer components of the Bellendir and Rass buffers are listed below the respective oligos. (B) A bar graph showing the first order rates of cleavage on either Bellendir substrates (5΄ flap-B and HJ-B) or Rass substrates (5΄ flap-R and HJ-R) in either the Bellendir Buffer or Rass Buffer by DmGen (at 25°C) or HsGEN1 (at 37°C). Colors for each substrate correspond to (A). Note that the final two bars are from experiments done in the same conditions of previously published data in (9). Each rate was determined from the average of ≥3 time courses (0 s–10 min or 30 min). Error bars are for the standard deviation of each mean rate. For rate values, see Supplementary Table S3.