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=Polyacrylamide Gel in tube Test= *[[Matt:LabNotes/2016-2-9 | Yesterday]] failed to get good complete polymerization even in the tubes ==Purpose== *Test polymerization of two different acrylaide:bis-acrylamide mixes (28:1 vs 199:1) *Test mineral oil as a sealant from oxygen ==Experiment== ===Samples=== #28:1, mineral oil, open lid #28:1, open lid #28:1, closed lid #199:1, mineral oil, open lid #199:1, open lid #199:1, closed lid ===Procedure=== *Following this general [[Matt:LabNotes/Polyacrylamide_Gel_Protocol]] *28:1 mix is made from 9 parts 29:1 and 1 part 19:1 *199:1 mix is made from 9 parts 40% acrylamide (from powder) and 1 part 19:1 *Added just a couple drops of mineral oil to cover **Good layer separation *Mix APS in PCR tubes and let sit on bench for 30min ==Results/Conclusion== *28:1 had noticeably stronger polymerization than 199:1 *After aspirating mineral oil good gel formed beneath *Open tube also has a polymerized gel but some liquid layer on top and sides that did not polymerize **Probably due to oxygen exposure *Overall open, closed, and mineral oil are all pretty comparable =Polyacrylamide Gel on culture dish= ==2 Conditions== #Try polymerizing in a vacuum chamber to minimize oxygen #*Stiff/dense gel formed along sides #Try polymerizing in a styrofoam box with CO2 from dry ice to displace oxygen ==Vacuum Chamber Procedure== *[[Matt:LabNotes/2016-1-27|Sample A1]] *Prepare 100ul 28:1 Acrylamide mix and add to dish *Put in vacuum chamber, apply vacuum, and then seal *Leave on bench for 30min ===Results=== *Stiff/dense gel formed along edges of hole *Center of glass has nothing *Guessing that vacuum caused water to evaporate ==Dry Ice Procedure== *[[Matt:LabNotes/2016-1-27|Sample A2]] *Prepare 100ul 28:1 Acrylamide mix and add to dish *Put in styrofoam box with beaker of water and drop dry ice in the water **Leave lid of box slightly ajar *After 30min no noticeable polymerization so kept it in the box with dry ice for 4 more hours ===Results=== *After ~5 hours the center was dry *The solution on edges was more viscous than before but not exactly a gel ==Conclusion== *Droplets formed after washing with 1X PBS that looked similar to "oil droplets" from mineral oil but there was never any mineral oil added **Possible that free radicals from APS or TEMED caused a small amount of [https://en.wikipedia.org/wiki/Elimination_reaction_of_free_radicals depolymerization of polystyrene]
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