Daniel:Notebook/PosSequencing/2017-4-10: Difference between revisions
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>Djacobse m (→Protocol) |
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<li>Repeat mixing to 100 uL (10 times)</li> | <li>Repeat mixing to 100 uL (10 times)</li> | ||
<li>Mix with vortexer for 3 minutes</li> | <li>Mix with vortexer for 3 minutes</li> | ||
<li>Dilute with pure mineral oil for imaging (1:200) | <li>Dilute with pure mineral oil for imaging (1:200)</li> | ||
</ol> | </ol> | ||
</ol> | </ol> |
Latest revision as of 16:34, 11 April 2017
Shao et al Droplet Making[edit]
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[edit]
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
- Dilute with pure mineral oil for imaging (1:200)
- 20170410-EmulsionTest2.jpg
Field 1
- 20170410-EmulsionTest3.jpg
Field 2
This mostly just serves as practice.