Daniel:Notebook/ComboLock/2016-3-15

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10% Acrylamide VA-044

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From yesterday's results we decided to increase the acrylamide concentration in the gel solution in order to make up for a lack of formaldehyde/tissue.

VA-044/10% Acrylamide test

For test 2 we used the same setup as test 1, but with the following recipe for the VA-044 gel mix:

Reagent Stock Concentration Final Concentration Amount added
Acrylamide 40% 9% 6.75 mL
Acrylamide:Bis (19:1) 40% (38:2) 0.95:0.05 (%) 0.75 mL
VA-044 ----- 0.25% (m/v) 0.075 g
PBS 10X 1X 3 mL
ddH2O ----- ----- 9.5 mL
Total     30 mL

This is a slight modification from yesterday's recipe. The idea is to be able to incorporate gel loading dye along with the solution. This will hopefully allow better visualization of droplets until we get the dyes we ordered. Also, the volume has been scaled down to 30 mL, and prior to loading in the syringe, loading dye and solution should be mixed 1:2.

Run Parameters

  • Line 1: Oil (on ice); 12 psi; ~2 mL
  • Line 2: Aqueous (VA-044P; on ice); 2.5 psi; ~1 mL

Results

By the end of the day, we had droplets but we still hadn't seen any signs of polymerization. As a test, I took an aliquot of our 10% acrylamide solution and put it in a 50 mL falcon tube. I then degassed the sample and added Argon before resealing the tube and leaving it at 37C. We also left the samples from the microfluidic experiments in the 37C incubator overnight (although they should need 3 hours or less, it was the end of the day).

In the morning I checked, and the in-tube reaction had indeed polymerized into a solid gel, indicating that the formula does work on the macroscale. On the microscale: