FAQ: Is it possible to reduce the volume of beads added during the second strand synthesis cleanup step?

The total volume of the sample when beads are added during the second strand synthesis cleanup step is 224 µl, which may exceed the volume of some 0.2 ml PCR tubes and plates. In addition this volume may not allow for complete separation of the beads using some magnets. Many PCR plates and tubes with a nominal value of 0.2 ml have a larger actual capacity that will accommodate a 224 µl sample. However, if the plate or tube you prefer to use does not allow for a 224 µl sample volume it is possible to reduce the volume of beads to 1.5 x (120 µl) for a total liquid volume of only 200 µl. In our hands we did not see a reduction in final library yield.