Hi, If we don't have Density gradient fractionation system ,...

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emma wang
Jun 13, 2019
protocol Protocol: Polysome Fractionation to Analyze mRNA Distribution Profiles
Hi, If we don't have Density gradient fractionation system , is there any other way to replace it? Is it feasible to separate gradient fractionation by SDS-PAGE? Do you have any suggestion? Thank you, Emma.
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Myriam Gorospe Author Answered Jun 18, 2019

Laboratory of Genetics and Genomics, National Institute on Aging, National Institutes of Health, USA

Following the sucrose gradient fractionation, it is possible to analyze specific proteins in each fraction by loading equal amounts of each fraction on SDS-PAGE then western blotting.
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Amaresh Chandra Panda Author Answered Jun 13, 2019

Institute of Life Sciences Bhubaneswar

The other option to collect the fractions by taking 1ml each from the top of the gradient manually. Here you can analyze your RNA in each fraction without having the 254nm RNA-distribution graph. Without that graph it's hard to know if everything worked well. I don't know if you can use SDS-PAGE for this.
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