Photopolymerization of acrylamide
July 7, 2005 10:15 AM
Attention molecular biology nerds: have you any experience with photoinitiation of acrylamide gel polymerization?
Our lab has an MJ BaseStation, which we purchased along with a ton of pre-packaged acrylamide, the kind that comes in an "instant ice-pack" style pouch. Said acrylamide includes a photoinitiator that lets us cast and polymerize a sequencing gel in less than 10 minutes. Very convenient, no need for APS/TEMED and long pre-runs on the sequencing machine.
Problem is, the pre-packaged stuff is getting old (4 years past the expiry date), and we're getting increased background noise compared to a fresh acrylamide that we polymerize the old-fashioned way. So, I'm looking to combine the best of both worlds -- fresh acrylamide that can be polymerized with a photoinitiator. Of course, we can buy a pre-made photoinitiator solution, but it's a bit pricey, and they won't tell us what's in it.
I've tried riboflavin, but it didn't work. This patent claims that 1-hydroxy-cyclohexyl-phenyl-ketone works well as a photoinitiator. The only problem I can see is that its max UV absorbance is at 250nm or so, while our UV box has its peak output at 312nm. For that reason, I thought 2-Methyl-4'-(methylthio)-2-morpholinopropiophenone might be a better choice.
So, to make a long story short, has anyone used either of these for photopolymerization? Or have you used anything else that works reliably?
Our lab has an MJ BaseStation, which we purchased along with a ton of pre-packaged acrylamide, the kind that comes in an "instant ice-pack" style pouch. Said acrylamide includes a photoinitiator that lets us cast and polymerize a sequencing gel in less than 10 minutes. Very convenient, no need for APS/TEMED and long pre-runs on the sequencing machine.
Problem is, the pre-packaged stuff is getting old (4 years past the expiry date), and we're getting increased background noise compared to a fresh acrylamide that we polymerize the old-fashioned way. So, I'm looking to combine the best of both worlds -- fresh acrylamide that can be polymerized with a photoinitiator. Of course, we can buy a pre-made photoinitiator solution, but it's a bit pricey, and they won't tell us what's in it.
I've tried riboflavin, but it didn't work. This patent claims that 1-hydroxy-cyclohexyl-phenyl-ketone works well as a photoinitiator. The only problem I can see is that its max UV absorbance is at 250nm or so, while our UV box has its peak output at 312nm. For that reason, I thought 2-Methyl-4'-(methylthio)-2-morpholinopropiophenone might be a better choice.
So, to make a long story short, has anyone used either of these for photopolymerization? Or have you used anything else that works reliably?
Thanks! Any idea at what concentration you used it?
posted by greatgefilte at 9:18 PM on July 7, 2005
posted by greatgefilte at 9:18 PM on July 7, 2005
This thread is closed to new comments.
I requested a free sample, and they sent me a whole bunch....
posted by mr_roboto at 12:33 PM on July 7, 2005