Daniel:Notebook/PosSequencing/2017-4-10

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Revision as of 16:06, 11 April 2017 by >Djacobse (→‎Protocol)
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Shao et al Droplet Making

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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.

  1. Oil phase
    1. Mix 5 uL Triton X, 10 uL Tween-20, and 450 uL Span 80 into 9.55 mL Mineral oil
    2. Vortex vigorously to mix
  2. Oil/aqueous mixing (adopted from Shao/Church)
    1. Mix 10 uL aqueous into 200 uL oil mixture
    2. Vortex for 30 seconds
    3. Repeat mixing to 100 uL (10 times)
    4. Mix with vortexer for 3 minutes
    5. Dilute with pure mineral oil for imaging (1:200)

This mostly just serves as practice.