Figure 3.
Robotic microdialysis set-up for assembly of recombinant RNAPs. (a) The photograph shows the placement of the 96-well microdialysis device on the robotic platform. A magnetic stirrer is sunk into the platform with full manual access to the stir speed control dial. The lid of the microdialysis device is held by the gripper arm of the robot during the loading and removal of the assembly reactions. On the left hand side, the peristaltic pump controlling the flow/dilution rate of the dialysis buffer can be seen. (b) Calculated urea concentration of the dilution of 200 ml 6M urea-containing buffer with urea-free buffer at a flow of 1 ml/min. After 17 h, the urea concentration in the dialysis buffer has effectively fallen to zero. The change of urea concentration (dc) over time (dt) can be expressed as a differential function (dc/dt) that is equal to the dilution (-D) of the initial concentration (c0) of urea according to the following equation ct = c0 × e −Dxt [D, Dilution (i.e. flowrate/volume); ct, urea concentration at time t; e, Euler's constant].