Subtractive cDNA cloning using magnetic beads and PCR.

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Subtractive cloning is a method that facilitates the isolation of nucleotide sequences present in a test sample but absent or present at much lower levels, in a reference sample. A variety of situations lend themselves to this methodology and it has been used, for example, to characterize changes in levels of gene expression during development (, , ), in pathological situations (, , ), in inductive events (,), and for the isolation of tissue-specific genes (,, , ). Current methodologies also allow detection of pathogens and integrated retroviruses present at exceedingly low levels in the cell ().