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The separation of RNA on the basis of size by sucrose gradient fractionation is a technique frequently employed in a cloning strategy (
1
–
3
). After production of poly(A)
+
mRNA by affinity chromatography (
see
Chapter 16 ) the RNA can be fractionated once or twice on sucrose gradients to produce a subpopulation of mRNA enriched for a particular species. The RNA fractions from the gradient can be assayed by in vitro translation and subsequent immunoprecipitation or bioassay of the products (
see
Chapters 19 –Chapters 22 ).