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=10% Acrylamide VA-044= [[Daniel:Notebook/ComboLock|Back to Calendar]] From [[Daniel:Notebook/ComboLock/2016-3-14|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: {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> |- style="background-color:#92CDDC;font-size:12pt;font-weight:bold" align="center" | width="142" height="30" | Reagent | width="98" | Stock Concentration | width="99" | Final Concentration | width="139" | Amount added |- style="font-size:12pt" | height="15" valign="bottom" | Acrylamide | align="center" align="center" valign="bottom" | 40% | align="center" align="center" valign="bottom" | 9% | align="center" valign="bottom" | 6.75 mL |- style="background-color:#D9D9D9;font-size:12pt" | height="15" valign="bottom" | Acrylamide:Bis (19:1) | align="center" valign="bottom" | 40% (38:2) | align="center" valign="bottom" | 0.95:0.05 (%) | align="center" valign="bottom" | 0.75 mL |- style="font-size:12pt" | height="15" valign="bottom" | VA-044 | align="center" valign="bottom" | ----- | align="center" valign="bottom" | 0.25% (m/v) | align="center" valign="bottom" | 0.075 g |- style="background-color:#D9D9D9;font-size:12pt" | height="15" valign="bottom" | PBS | align="center" valign="bottom" | 10X | align="center" valign="bottom" | 1X | align="center" valign="bottom" | 3 mL |- style="background-color:#D9D9D9;font-size:12pt" | height="15" valign="bottom" | ddH2O | align="center" valign="bottom" | ----- | align="center" valign="bottom" | ----- | align="center" valign="bottom" | 9.5 mL |- style="font-size:12pt" |style="font-weight:bold" height="15" valign="bottom" | Total | align="center" align="center" valign="bottom" | | align="center" align="center" valign="bottom" | |style="font-weight:bold" align="center" valign="bottom" | 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: <gallery heights=400px widths=400px perrow=2 caption="Results from Drop-Seq Device Test 03/15/16"> File:20160316_A3_A4_Fractions.jpg|Fractions 3 and 4 from Drop Seq device test after overnight incubation at 37C. Notice that both fractions contain an oil phase (transparent) above an aqueous phase (blue) File:20160316_A3A4_Fractions_Inverted.jpg|Fractions 3 and 4 from Drop Seq device test after overnight at 37C, inverted. Notice that the aqueous phase remains stuck to the top of the tube, whereas the oil phase has dropped to the bottom. </gallery>
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