High Throughput Sequencing (HTS)
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The CGRB began offering the Illumina/Solexa high throughput DNA sequencing service in April 2007. The paired end module was added June 2008. In January 2009, the Illumina 1G Genome Analyzer was upgraded to the GAII, which allows for increased data output, longer reads, improved accuracy, and faster run times. The system is operational and ready for a variety of applications. The sequencing process takes place on a flow cell with 8 channels (one channel must be used for control DNA), each of which may contain a different sample. Over 360 Mb of raw sequence data are generated per channel for a single end 36 cycle run, although usable, high-quality data will be less. For expression profiling, up to 10 million tags are sequenced per channel. |
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Illumina sells different sample prep kits depending on the application: Genomic DNA, NlaIII Gene Expression, DpnII Gene Expression, Small RNA, Paired End Genomic DNA, Mate Pair Library, ChIP-Seq, mRNA-Seq, and Multiplex. Sample prep reagents should be purchased directly from Illumina. Please contact Mark Dasenko for more information and assistance with getting started. Additionally, please be well aware of the quantity of image and sequence data that will be generated during a run (nearly 1 Tb). This will require significant computational power and expertise. The CGRB will provide a number of service, software and hardware tools to facilitate capture and analysis of data. However, each group should consider their long-term computational programming and utilization needs, and plan accordingly with personnel. To help the Illumina 1G user community, we established an HTS Group that meets bi-weekly and shares information. Please contact Scott Givan for more information and recommendations, or to join the HTS group.
An overview of the 1G workflow and approximate durations of steps for a typical |
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