Matt:LabNotes/2016-2-10: Difference between revisions
Jump to navigation
Jump to search
>Mzcai (Created page with "=Polyacrylamide Gel in tube Test= * Yesterday failed to get good complete polymerization even in the tubes ==Purpose== *Test polymerization of two...") |
>Mzcai m (→Conclusion) |
||
Line 59: | Line 59: | ||
==Conclusion== | ==Conclusion== | ||
*Droplets formed after washing with 1X PBS that looked similar to "oil droplets" from mineral oil but there was | *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] |
Latest revision as of 20:09, 12 February 2016
Polyacrylamide Gel in tube Test[edit]
- Yesterday failed to get good complete polymerization even in the tubes
Purpose[edit]
- Test polymerization of two different acrylaide:bis-acrylamide mixes (28:1 vs 199:1)
- Test mineral oil as a sealant from oxygen
Experiment[edit]
Samples[edit]
- 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[edit]
- 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[edit]
- 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[edit]
2 Conditions[edit]
- 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[edit]
- 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[edit]
- Stiff/dense gel formed along edges of hole
- Center of glass has nothing
- Guessing that vacuum caused water to evaporate
Dry Ice Procedure[edit]
- 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[edit]
- After ~5 hours the center was dry
- The solution on edges was more viscous than before but not exactly a gel
Conclusion[edit]
- 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 depolymerization of polystyrene