Makoto Kawase，the National Institute of Agrobiological Sciences in Japan
http://www.geocities.jp/millet_kawase/rlcs/RLCS_protocol.html
This protocol is for inmature barley grains，probably applicable for other plant materials
RLCS that was developed by Suzuki et al. (1996) displays many cDNA species quantitatively and simultaneously at two dimensional gel spots (RLCS profile). In this method, cDNA species of uniform length were prepared for each mRNA species using restriction enzymes. After the restriction, enzyme sites were radiolabeled as landmarks, and then the labeled fragments were subjected to high resolution two-dimensional gel electrphoresis. Since the location and intensities of spots reflect the expression of certain mRNA species, RLCS serves as a usuful cDNA display system that provides valuale information on expressed genes.
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