Project description:The aim of this experiment was to compare the transciptome of the peach-potato aphid (Myzus persicae) clone 4106a (a laboratory insecticide-susceptible standard collected from potato in Scotland in 2000) with clone FRC (an insecticide resistant aphid clone collected from peach in France in 2009) to identify which genes are over or underexpressed in the resistant phenotype. The custom microarray used in this study was designed using the Agilent eArray platform (Agilent Technologies) by the Georg Jander Lab and is based on a previously described array containing probes for >10, 000 M. persicae unigenes produced by Sanger sequencing (Ramsey, Wilson et al. 2007) augmented with an additional 30, 517 probe set designed on EST unigene sequences identified in a 454 sequencing project (Ramsey, Rider et al. 2010). The final slide layout consists of four arrays of 45, 220 60-mer probes and these are produced by Agilent by in situ oligonucleotide synthesis. References: Ramsey, J. S., D. S. Rider, et al. (2010). "Comparative analysis of detoxification enzymes in Acyrthosiphon pisum and Myzus persicae." Insect Molecular Biology 19: 155-164. Ramsey, J. S., A. C. C. Wilson, et al. (2007). "Genomic resources for Myzus persicae: EST sequencing, SNP identification, and microarray design." BMC Genomics 8.
Project description:The aim of this experiment was to compare the transciptome of the peach-potato aphid (Myzus persicae) clone 4106a (a laboratory insecticide-susceptible standard collected from potato in Scotland in 2000) with clone FRC (an insecticide resistant aphid clone collected from peach in France in 2009) to identify which genes are over or underexpressed in the resistant phenotype. The custom microarray used in this study was designed using the Agilent eArray platform (Agilent Technologies) by the Georg Jander Lab and is based on a previously described array containing probes for >10, 000 M. persicae unigenes produced by Sanger sequencing (Ramsey, Wilson et al. 2007) augmented with an additional 30, 517 probe set designed on EST unigene sequences identified in a 454 sequencing project (Ramsey, Rider et al. 2010). The final slide layout consists of four arrays of 45, 220 60-mer probes and these are produced by Agilent by in situ oligonucleotide synthesis. References: Ramsey, J. S., D. S. Rider, et al. (2010). "Comparative analysis of detoxification enzymes in Acyrthosiphon pisum and Myzus persicae." Insect Molecular Biology 19: 155-164. Ramsey, J. S., A. C. C. Wilson, et al. (2007). "Genomic resources for Myzus persicae: EST sequencing, SNP identification, and microarray design." BMC Genomics 8. Two-condition experiment, 4106a vs. FRC Myzus persicae clones. Biological replicates: 4 pools of RNA extracted from ten 15 day old aphids of each clone. Technical Replicates: Two technical reps incorporating a dye swap. Total replication: eight replicates for each clone.
Project description:Background: Understanding the genetic elements that contribute to key aspects of coffee biology will impact future agronomical improvements for this economically important tree. The past years, EST collections were generated in Coffee, opening the possibility to create new tools for functional genomics. Results: The project PUCE CAFE, set up by the scientific consortium NESTLE/IRD/CIRAD has developed of long oligonucleotide coffee array using public coffee EST sequences mainly obtained from different stages during fruit development and leaves in Coffea canephora (Robusta). We have performed a validation experiment in order to check the array usability and the reproducibility of hybridizations. Conclusion: We have generated the first 15K Coffee array during this three years project PUCE CAFE, granted by The French National Research Agency (ANR, Programme Génoplante) . This new tool was dedicated to large scale transcriptomic analysis during grain development of Coffea canephora grown in different countries . Furthermore, other analysis have been also initiated by the different partners like analysis of polyploidy or drought resistance. In any case, at the end of the project, the generated arrays will be available to the international scientific community.