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==Protocol Part 2== <ol start="6"> <li>Emulsion Breaking</li> <ol type="A"> <li>With a 200 uL pipette pool the emulsion into a 1.5 mL tube; save the tip and put aside</li> <li>Add 100 uL isopropyl alcohol (IPA) to tube with 200 uL tip (use new tip)</li> <li>Draw up IPA with tip from part A, mix to resuspend tip</li> <li>Vortex the pooled sample in the 1.5 mL tube for 30 seconds</li> <li>Spin down the sample 13,000 rpm for 5 minutes</li> <li>Take off oil-IPA mix (top layer) but do not disrupt the oil/aqueous interface</li> <li>Add 1000 uL IPA and vortex for 30 seconds</li> <li>Spin down sample at 13,000rpm for 3 minutes</li> <li>Again remove IPA without disturbing the pellet</li> <li>Add 500 uL 4X SSC wash buffer to 1.5 mL tube and vortex for 30 seconds</li> <li>Go back and mix thoroughly by pipetting up and down; the pellet may be hard to resuspend</li> <li>Leave on 1.5mL mixer at 1300rpm 1 minute</li> <li>Spin down sample at 13,000rpm for 1 minute</li> <li>Remove all but 40uL wash buffer</li> <li>Repeat steps 11 to 15 two times (j to o)</li> <li>Resuspend in ~100 uL 4X SSC buffer</li></ol> <li>Fluorescent Oligo Binding</li> <li></li> <li>Imaging</li> </ol>
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