Hey,
I was wondering if someone has prepared linear polyacrylamide (LPA) themselves before? I wanted to use it in DNA precipitation but it was far too expensive to buy whereas we already have acrylamide in the lab.
My main question is whether I could filter the solution or not? To make sure it is very clean since it needs to be used with DNA and later PCR. But of course LPA is a polymer so I fear it would not pass through the filter.
not an answer to your question but I have often found glycogen is effective and easy to use as a co precipitant polymer that has little effect on downstream applications
Dear all,
there are a lot of references on the preparation of LPA. The attached file is an exemple. Regards
I went with LPA because my project involves aptamers and there could be a chance the DNA will bind to glycogen for example. Especially, since I'm working with 66bp DNA I thought LPA would have the best chance for high recovery of so small sized DNA.
I did use TEMED for polymerisation. It is quite diluted I think 5mg/mL. I suppose I can just try to filter it and think if it works or not. My first attempt of ethanol precipitation with and without LPA didn't show any difference in the concentration of the DNA in an agarose gel. However, it did help me visualise the pellet.
I use the recipe given by Nadia except that mix only acrylamide, TEMED and APS (without Tris, EDTA and salt). Filter the solution through 0.22um syringe filter before adding APS (you can prepare 5% extra solution to compensate for the leftover volume blocked in filter).
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