Biotechnology and Bioinformatics (1+1)

Random primer labeling of DNA

The Klenow fragment of E. coli DNA polymerase I carries the polymerase and 3’→5’ exonuclease activities but lacks the 5’→ 3’ exonuclease activity. This method originally developed by Feinberg and Vogelstein (1983) employs its 5’→ 3’ polymerase activity. Synthetic random primers of 6-10 bases are used as primers which form hybrids at many positions along the template. The template for labeling must be denatured prior to the reaction.