Analysis of the size distribution of first-strand cDNA molecules.

Abstract

We have developed a method to analyze the size distribution of the first-strand cDNA molecules corresponding to given mRNA species. First-strand molecules synthesized from cytoplasmic polyadenylated RNAs are separated by electrophoresis on an alkaline agarose gel, and a Southern blot hybridization is performed. As an example, we analyzed the first-strand molecules corresponding to the human c-myc mRNAs. This method can be used to determine whether full-length, first-strand molecules corresponding to an mRNA species to be cloned are synthesized efficiently. Interestingly, this method allows one to analyze full-length, first-strand cDNA molecules with a much higher resolution than Northern blot analysis of mRNA molecules. This method can therefore be used to discriminate between the multiple mRNA species transcribed from a given gene or the homologous mRNA species transcribed from a given gene family.