Improvement of the print-capture polymerase chain reaction procedure for efficient amplification of DNA virus genomes from plants and insect vectors

A rapid and simple procedure is described to amplify efficiently geminiviru s DNA genomes by improving the print-capture polymerase chain reaction (PCR ) procedure reported recently for RNA viruses. This method, termed print-PC R (P-PCR), allows direct amplification of DNA from infected plant or whitef ly tissues printed directly on Whatman 3MM paper, without the need of any g rinding, incubation, or washing steps previous to the amplification reactio n. P-PCR reduces sample manipulation and avoids previous extraction of nucl eic acids, thereby diminishing the possibilities of cross-contamination bet ween samples. P-PCR has been successfully applied to whiteflies and various plant species infected by two different tomato yellow leaf curl viruses, T YLCV-Sr and TYLCV-Is, and for the amplification of the full-length genome o f TYLCV-Is from infected plants. (C) 1998 Published by Elsevier Science B.V . All rights reserved.