cDNA Synthesis and Cloning

The isolation of intact messenger RNA and its conversion into cDNA copies by avian or Moloney murine reverse transcriptase, as well as subsequent amplification of gene transcripts by the PCR technique, are becoming increasingly important tools in molecular biology. At present, these techniques have been often necessary and widely used for the analysis of individual mRNA levels in cells and tissues by Northern blot analysis, nuclease protection analysis and in situ hybridization. Another important application of RNA templates is the construction of representa­tive cDNA libraries in order to clone genes, to investigate their molecular structure and to express them in prokaryotic and/or eukary…