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==Protocol== <ol start="2"> <li>Qiaquick Column</li> '''Note''': I will be pooling samples and double loading columns, yielding 4 samples per column (2 columns per barcode oligo, 12 columns total) <ol type="A"> <li>Pool 4 samples and 1000 uL PB (5X binding buffer)</li> <li>Load sample onto column and spin for 1 minute at 14000 rpm; discard flow through</li> <li>Add another 600 uL pooled sample to columnand spin for 1 minute at 14000 rpm; discard flow through <li>Add 700 uL PE (wash buffer) to column and spin for 1 minute at 14000 rpm; discard flow through</li> <li>Dry spin column for 1 minute at 14000 rpm; discard flow through</li> <li>Let stand with cover open in fume hood for ~5 minutes</li> <li>Transfer column to a new 1.5 mL eppendorf tube</li> <li>Add 35 uL nfH<sub>2</sub>O to column</li> <li>Let stand 1 minute</li> <li>Spin for 1 minute at 14000 rpm</li> <li>Pool samples when finished; concentrate if desired</li> </ol> <li>TBE Gel</li> <ol type="A"> <li>Mix 160 uL TBE and 40 uL 6X dye</li> <li>Aliquot 10 uL mix to parafilm</li> <li>Add 2 uL sample or 1.5 uL ladder to appropriate aliquot</li> <li>Add 10 uL mix to appropriate lane</li> <li>Run gel at 240V for 24 minutes</li> <li>Open gel and stain with 2 uL SYBR gold for 3 minutes</li> <li>Rinse and image in gel doc</li> </ol> <li>Qubit</li> <ol type="A"> <li>Mix 1194 uL dsDNA buffer and 6 uL dye</li> <li>Aliquot 190 uL to standard tubes and 199 uL to dye tubes</li> <li>Add 10 uL standard or 1 uL dye</li> <li>Vortex and spin down</li> <li>Let sit 5 minutes in the dark</li> <li>Measure in the Qubit</li> </ol></ol>
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