In an outbreak situation in crops, it is necessary to obtain quickly a good overview of the viruses that may be involved. Only in this way can the right measures be taken to safeguard crop production immediately but also in the long term. High-throughput sequencing (HTS) and bioinformatics analysis of samples offer a global analysis of the phytosanitary status of plants and the rapid and precise identification of pathogens.
Unit: Analysis of ten individual samples in an outbreak situation, covering all steps of a workflow - from DNA/RNA extraction, state of the art library preparation, sequencing on Illumina platforms and comprehensive bioinformatics analysis.
Aeonium ringspot virus, RNA1 and RNA2 full-length cDNAs are cloned in plasmid pUC18, under the control of a T7 promoter, allowing for transcription of infectious RNAs.
Unit: 2 micrograms of each plasmid DNA, allowing for the inoculation of six host plants (Nicotiana benthamiana) after T7-driven transcription
The 3' terminal 217 nts of Aeonium ringspot virus RNA1 and RNA2 (excluding the poly(A) tail) are cloned in pGEM-T Easy allowing for transcription of a riboprobe to detect both genomic RNAs.
Unit: 2 micrograms of plasmid DNA, allowing for the synthesis of a probe after T7-driven transcription