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=== Step 4. Washing the Capture Beads Plus Bound DNA === # After 45-min incubation, add 100ul of 1X Wash Buffer I heated to 47C to the 15ul capture beads plus bound DNA. I added while the tube maintained on thermocycler at 47C. # Mix by vortexing 10X # Transfer the entire contents to Lo-bind 1.5mL tube # Place the tube in the magnet to bind the beads. Remove and discard spnt # Remove the tube from the magnet and add 200ul of 1X Stringent Wash Buffer heated to 47C. Pipette up and down 10X. WORK QUICKLY so that the temperature does not drop down much below 47C. ## I place the tube to thermomixter next to Stringent Wash Buffer and add the buffer. # Incubate at 47C for 5min # Repeat step 4.4 - 4.6 for a total 2 washes using 1X stringent Wash Buffer heated to 47C # Place the tube back to the magnet and remove spnt # Add 200ul of RT 1X Wash Buffer I -> mix by vortexing for '''2min''' # Place the tube back to the magnet and remove spnt # Add 200ul of RT 1X Wash Buffer II -> mix by vortexing for '''1min''' # Place the tube back to the magnet and remove spnt # Add 200ul of RT 1X Wash Buffer III -> mix by vortexing for '''30s''' # Place the tube back to the magnet and remove spnt # Remove the tube from the magnet and add 50ul H2O. Mix by vortexing. # Continue to Amplification Step or store the beads plus captured DNA at -15 to -20C. # I stored the captured DNA in -20C '''THERE IS NO NEED TO ELUTE DNA FROM THE BEADS'''<br> <br>
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