HOME   PERSONNEL   PUBS   FIELD CAMPAIGNS   COURSES   DIAGNOSTIC TOOLS   LINKS   MEMBERS ONLY   << back to Research PCR Cleanup We use primarily ExoSAP-IT© for PCR Product Clean-Up for standard cleanup of successful PCR. Alternatively we occasionally use a gel purification and isolation technique in which case we use the Gene Clean method Run standard gel (~1.5%) with 1X TAE (TBE does not work as well) Cut out band using clean razor blade Weigh agarose in mg Add 3X agarose weight in microliters of NaI Incubate 5-10 min at 55oC until agarose is thoroughly dissolved Suspend Glass Milk by thorough vortexing Add 5 l Glass Milk to agarose solution Mix by inverting and vortexing Place on ice for 5 min vortexing about every 2 min Centrifuge tube on high 5 sec Pipette off supernatant Keep tubes on ice as much as possible. Add 600 l of cold New Wash, and resuspend pellet by vortexing. (note: store New Wash tightly sealed at -20ºC) Centrifuge tube on high 15 sec Pipette off supernatant Go to step 12 two more times Air dry for 15-20 min Resusped in 12 l TE buffer or ultra-pure H2O Incubate ate 45 -55oC for 3 min Centrifuge 30 sec Remove supernatant (with DNA) into clean tube

Kelly B. Miller Lab, Department of Biology, University of New Mexico, 167 Castetter Hall, MSC03 2020, Albuquerque, NM 87131-0001 USA Contact the website coordinator with questions or comments regarding this website.