Improved RAPD amplification of recalcitrant plant DNA by the use of activated charcoal during DNA extraction

Abstract

Abstract An efficient procedure has been developed for DNA extraction from cotton by modifying the original CTAB method. The major improvement concerns the use of activated charcoal to bind resinous and coloured compounds which copurify with the DNA. The efficiency of amplification by RAPD was used as a criterion to evaluate the action of activated charcoal. Twenty‐five random decamers from Operon Technologies were used to compare DNA samples extracted in the presence or absence of activated charcoal. The results in terms of amplification suggest that the use of activated charcoal in DNA extraction enhances RAPD amplification. This technique was initially developed for cotton but it has been applied successfully to other recalcitrant plants such as coffee, rubber tree, cassava and banana.