New device and method for capture, reverse transcription and nested PCR in a single closed-tube.
A device and improved method based on the use of a compartmentalized Eppendorf tube that allows capture, reverse transcription and nested-PCR in a single closed-tube has been developed and patented. The main advantages of the system are the high sensitivity obtained, the simplicity, the low risk of contamination and the easy establishment of adequate conditions for nested-PCR. The method has been successfully applied to the detection and characterization of citrus tristeza closterovirus and plum pox potyvirus isolates in plant tissues and single aphids squashed on paper. This device and methodology could be easily adapted to the detection of other targets.
Departamento de Protección Vegetal y Biotecnología, Instituto Valenciano de Investigaciones Agrarias (IVIA),Apartado oficial, 46113 Moncada, Valencia, Spain.
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Olmos
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Cambra
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Esteban
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Gorris
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Terrada
D000818:Animals; D001042:Aphids / Q000821:virology*; D017790:Closterovirus / Q000302:isolation & purification*; D017801:Plum Pox Virus / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000295:instrumentation*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000295:instrumentation*