Sanger Sequencing and Tilling:

The DNA sequencing service provides high-quality Sanger sequencing of various templates on a routine low-scale (single-tube) and large-scale (96- and 384-well plates) basis. We provide quick turnaround times, and professional support to guarantee excellent data quality. The TILLING (Targeting Induced Local Lesions IN Genomes) project was initiated as collaborative effort of several research groups in the MPI CBG and the Biotec to provide a tool to screen for point mutations or sequence variants in any gene of interest. This technology is based on large-scale amplification of a gene fragment of interest by PCR from libraries of mutagenized individuals and subsequent Sanger sequencing to detect sequence variants. TILLING libraries are available for various model organisms such as the zebrafish D. rerio, the fruitfly Drosophila melanogaster, and the worm C. elegans. Modified TILLING approaches have been established to detect sequence variants naturally that occur in plant species such as the aspen.

Services:

• DNA sequencing (single tube and plate format)
• Primer walking projects
• High-throughput fosmid end sequencing
• Large-scale sequencing of PCR fragments
• Fragment analysis
• Generation and handling of TILLING libraries in zebrafish
• Design and establishment of TILLING projects in zebrafish and fly
• EcoTILLING in plant species

Expertise:

• High-quality DNA sequencing and professional support
• Extensive experience in zebrafish genetics and genomics
• Liquid handling for PCR- and Sanger sequencing reactions
• Assay development for genotyping projects (variation detection)