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==Protocol (Part 3)== Since the TBE gel showed a smear at the correct range (104 bp, but with a lot of DNA and biotin attached it runs funny), I'll be moving on to a size select and purification. <ol start="7"> <li>Size select gel</li> <ol type="A"> <li>Mix 40 uL TBE, 10 uL loading dye, and 10 uL sample for C1-C2 and C3-C2</li> <li>Mix 23 uL TBE, 5 uL loading dye, and 2 uL ladder</li> <li>Add 30 uL to each lane (2 lanes for C1-C2 and C3-C2)</li> <li>Run the gel at 250V for 25 min</li> <li>Stain with 3 uL SYBR gold for 3 min</li> <li>Image, and extract bands between 100bp and 125 bp; collect the gel bands in the same tube</li> <gallery perrow=2 heights=200px widths=200px> File:2016-08-16-PosControlAmplicon-SizeSelect.png|Before Image File:2016-08-16-PosControlAmplicon-SizeSelect-After.png|After Image </gallery> <li>Shred the gel by centrifuging at 14000rpm for 1 min 30 sec</li> </ol> As you can see from the gel images, there was too much DNA on the gel and it ran funny. Besides, the caps fell off in the centrifuge. [[Category:ComboLock]] [[Category:20160805]]
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