DNA sequencing is performed using the Applied Biosystems Automated 3730 DNA Analyzer. We use Big Dye Terminator chemistry and AmpliTaq-FS DNA Polymerase. We routinely provide up to 900 bases per reaction, providing the template is of high quality. Other services associated with DNA sequencing are also offered. Our hours are 8 am to 4:30pm, Monday through Thursday and 8am-4pm Friday. We can be reached at (607) 254-4857.

DNA Sequencing is now available using an Illumina/Solexa Genome Analyzer. This technology can be used for genomic resequencing projects, targeted resequencing projects, small RNA discovery and analysis, sequencing of DNA fragments isolated by chromosome immunoprecipitation (ChIP-Seq), and Digital Gene Expression analysis. These services can be arranged by consultation with the Director of the DNA Sequencing and Genotyping Laboratory.

Our Applied BioSystems 3730xl DNA Analyzer has the ability to determine the size of your DNA fragments by using a fluorescence-based detection system. The use of five fluorescent dyes allows multiplexing of 4 microsatellites (for example) plus a size standard in every lane, eliminating lane to lane variability and increasing throughput. We accept samples in 96 or 384 well plates, and you must supply your own size standard. Please contact us for more information.

Applied Biosystems has developed GeneMapper Software v3.0, which combines the functions of GeneScan and Genotyper software in one convenient package. Designed to support a wide variety of research applications, GeneMapper software is ideal for SNP validation and screening, microsatellite analysis, and sizing-only applications. This software can analyze fragment analysis data from our 3730xl. When used with the recommended computer platform, GeneMapper Software v3.0 will process more than 50,000 genotypes in one hour.

Instructions on accessing Genemapper via Microsoft Remote Desktop

The Evolutionary Genetics Core Facility (EGCF) specializes in the construction and screening of genomic DNA libraries enriched for microsatellites, optimization of data collection and analysis, and project consultation. Also, the EGCF runs a microsatellite development course (BioEE 761) every fall semester. Contact Steve Bogdanowicz for additional information and details.

The DNA Sequencing and Genotying facility offers the use of Real Time PCR and Sequence Detection instrumentation. You can analyze your plates on an Applied Biosystems 7900 HT Sequence Detection System. This system allows quantitation of starting material (genomic DNA, plasmid, RNA) along with heterozygote detection and SNP genotyping.

The user is responsible for purchasing consumables and setting up the reactions in plates compatible with our instrument. Time may be reserved by signing up in 147 Biotech or by calling the lab at 254-4857.

Primer design and data analysis software are available to create primer and probe pairs specifically designed for your target sequence or to analyze your data. The instrument uses either 96- or 384-well plates and uses either fluorescent probes or the SYBR green reagent for detection. The charge is the same regardless of which plate you use.