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==Linear Product Production-Bglii (Started [[Daniel:Notebook/ComboLock/2017-3-25|Yesterday]])== <ol start="2"> <li>TBE gel</li> <ol type="A"> <li>Mix 64 uL TBE and 16 uL 6X dye</li> <li>Aliquot 10 uL on to parafilm per sample/ladder</li> <li>Add 2 uL reaction, 1 uL (10 uM) controls, and 1.5 uL ladder to appropriate aliquot</li> [[Image:GelLanes-20170325-LinearProduct.png|400px]] <li>Add 10 uL mix to gel lanes</li> <li>Run gel for 24 minutes at 230V</li> <li>Stain with 2 uL SYBR gold for 3 minutes</li> <li>Rinse and image in gel doc</li> </ol></ol> [[Image:2017-03-26-LinearProductProduction-Bglii.png|400px]] That's the right one. However, there is a ton of primer (bottom of the gel), so I'll run a ssDNA column to fix. <ol start="3"> <li>ssDNA column</li> <ol type="A"> <li>Add 50 uL '''Binding Buffer''' to the sample; mix well</li> <li>Transfer to '''IIC Column''' and centrifuge at 14000 rpm for 1 minute; '''SAVE THE FLOW THROUGH'''</li> <li>Add 70 uL 100% EtOH to flow through; mix well</li> <li>Transfer to '''IC Column''' and centrifuge at 14000 rpm for 1 minute; discard flow through</li> <li>Add 400 uL '''Prep Buffer''' and centrifuge at 14000 rpm for 1 minute; discard flow through</li> <li>Add 700 uL '''Wash Buffer''' and centrifuge at 14000 rpm for 1 minute; discard flow through</li> <li>Add 400 uL '''Wash Buffer''' and centrifuge at 14000 rpm for 1 minute; discard flow through</li> <li>Centrifuge empty column at 14000 rpm for 2 minutes</li> <li>Transfer to empty 1.5mL centrifuge tube (low bind)</li> <li>Add 20 uL nfH2O and centrifuge at 14000 rpm for 1 minute</li> </ol>
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