Matt:LabNotes/2016-2-10
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Polyacrylamide Gel in tube Test
- 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
- 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
- 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 no mineral oil