Daniel:Notebook/PosSequencing/2017-4-10
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Shao et al Droplet Making
Basically just messing around with this one, but I want to do this to start to get an idea of what to test for when making droplets. Hopefully if I focus carefully on the droplet making for now the qPCR will work better.
Protocol
0.05% Triton The basic protocol calls for 4.5% Span-80, 0.4% Tween-80, and 0.05% Triton X-100 in mineral oil. Since I don't actually have Tween 80, I'll use Tween -20 at 0.1%, which is high for other protocols I've used.
- Oil phase
- Mix 5 uL Triton X, 10 uL Tween-20, and 450 uL Span 80 into 9.55 mL Mineral oil
- Vortex vigorously to mix
- Oil/aqueous mixing (adopted from Shao/Church)
- Mix 10 uL aqueous into 200 uL oil mixture
- Vortex for 30 seconds
- Repeat mixing to 100 uL (10 times)
- Mix with vortexer for 3 minutes