Objectives

The aim of the Agap functional analysis platform is to provide plant scientists with the environment and tools to carry out studies on Mediterranean and tropical species that elucidate gene functions.

Each year, the platform hosts numerous projects. Most of them seek to determine the genetic and functional bases of plant development and how plants adapt to their environment. Thus, the genes studied on this platform are involved in the development, metabolism and response of plants to stress, mechanisms directly or indirectly involved in the agricultural value of a plant. They are generally identified beforehand thanks to comparative sequencing, or to methods based on association genetics or genome-wide expression studies, which are rapidly developing, but the AFEG platform enables them to be studied on a more focalized scale, thanks to multiple molecular biology techniques.

Know-how, tools and resources

The functional genomics platform has the equipment, tools and reagents needed to study the expression of genes from monocots and dicots, and the constitution of expression vectors.

The platform is divided into various workshops specializing in:

• Cloning sequences of interest (Gateway, restriction)
• Transformation of bacteria
• Amplification of fragments up to 15 kb
• Quantification and visualization of nucleic acids
• Extraction, visualization and production of proteins
• RNA extraction/cDNA synthesis
• Quantitative PCR
• Genome editing tools, such as CRISPR/Cas9

The members of the platform keep up to date with technological developments and provide hosting and training for people of various origins (students, technicians, researchers).

A training course "Appliquer la technologie CRISPR-Cas9 en plantes: de la théorie à la pratique" (Applying CRISPR-Cas9 technology to plants: turning theory into practice) is proposed by the AFEG platform, on request. The latest session can be consulted here.

Equipment and installations

The functional analysis platform is located at CIRAD’s Lavalette site. It is available to all the teams in the Unit and their partners.

It has:

• Assay devices: TapeStation (Agilent), Bioanalyser (Agilent), Nanoquant (Tecan), Fluoroskan Ascent (Thermo Scientific)
• 3 real time thermocyclers (LC480 from Roche and Biomark from Fluidigm),
• 2 thermocyclers including 1 gradient
• Numerous PCR blocks (minicycler, 96 well block),
• UV lamp + camera
• Migration tanks
• Bacteriological hoods (x2)
• 2 Biorad electroporators
• Climo Shaker ISFI-XX incubator
• Chemical hood
• Common laboratory equipment (Centrifuges, ice machine, freezers and refrigerators, vortex, water baths and dry baths, ovens, electronic pipettes)
• Freezer -80°