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 geminivirus DNA genomes by improving the print-capture polymerase chain reaction (PCR) procedure reported recently for RNA viruses. This method, termed print-PCR (P-PCR), allows direct amplification of DNA from infected plant or whitefly tissues printed directly on Whatman 3MM paper, without the need of any grinding, incubation, or washing steps previous to the amplification reaction. P-PCR reduces sample manipulation and avoids previous extraction of nucleic acids, thereby diminishing the possibilities of cross-contamination between samples. P-PCR has been successfully applied to whiteflies and various plant species infected by two different tomato yellow leaf curl viruses, TYLCV-Sr and TYLCV-Is, and for the amplification of the full-length genome of TYLCV-Is from infected plants.
Navas-Castillo
J
J
Estación Experimental La Mayora, CSIC, Málaga, Spain.
Díaz
J A
JA
Sánchez-Campos
S
S
Moriones
E
E
D000818:Animals; D004279:DNA, Viral / Q000032:analysis; D004587:Electrophoresis, Agar Gel; D017791:Geminiviridae / Q000235:genetics* / Q000302:isolation & purification; D016679:Genome, Viral; D006430:Hemiptera / Q000821:virology; D007303:Insect Vectors / Q000821:virology; D010944:Plants / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods*
D004279:DNA, Viral