Prep+for+electroelution+experiment+3-21-13

plan electroelution of DNA <> -dialysis membrane for electroelution of DNA -http://humgen.wustl.edu/hdk_lab_manual/rdm/rdm9.html --SPECTRA/POR dialysis tubing (VWR, B, F 10mm x 50 ft., cat.#132645) and clips ---this product corresponds to a molecular weight of 6000 to 8000

-D-Tube Electroelution Accessory Kit --they use Mini, Midi, or Maxi membranes with DNA (found in manual here: <> http://www.emdmillipore.com/life-science-research/d-tube-electroelution-accessory-kit/EMD_BIO-71511/p_C3Cb.s1OuKYAAAEjKBp9.zLX?attachments=USP

 -smallest cutoffs for Mini, Midi, and Maxi are 6, 3.5, and 3.5

-dialysis in our lab <> We have 1,000 MWCO tubing (Spectra/Por CE (Cellulose Ester) Membrane 131090 <> http://www.spectrumlabs.com/dialysis/BiotechTubing.html?Pn=131090;   It looks like this tubing should work for me

-basic notes about protocol (from here: http://humgen.wustl.edu/hdk_lab_manual/rdm/rdm9.html) -minimize exposure to UV (use prep (long form) of UV to cut out the DNA -Cut a piece of dialysis tubing approximately 3 cm longer than the gel slice and clip one end. Gently push gel slice into open end and down to the clip. Add some buffer -Place gel slices parallel to the electrodes and fill the electrophoresis box with buffer (again, same as original gel) until all tubing is submerged. -Electroelute at 80-100 volts for 2-5 hours, longer for large fragments. The movement of the DNA can be monitored with a hand held UV source (long-wavelength). -DNA may stick to dialysis tubing after current flow. Remove the gel piece from the dialysis tubing. Reverse the electrodes and run for 15 s to 2 min to dislodge the DNA from the membrane surface.

