From GenoSeq
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GS FLX+ System : High Throughput Sequencer
To find out more about using this technology in your research contact us.
Sanger-like read lengths - the power of next-gen throughput.
The complete sequencing workflow of the GS FLX and GS Junior Systems comprise of four main steps, leading from purified DNA to analyzed results. These basic steps include:
- Generation of a single-stranded template DNA library:
- Emulsion-based clonal amplification of the library:
- Data generation via sequencing-by-synthesis:
- Data analysis using different bioinformatics tools:
To find out more information about the technology and how the system works please click on the following link The Technology:
Sequencing Kits
| Sequencing Kits | GS FLX Titanium XL+ (Shotgun Sequencing) | GS FLX Titanium XLR70 (Shotgun & Amplicon Sequencing) |
|---|---|---|
| Read Length | Up to 1,000 bp | Up to 600bp |
| Mode Read Length | 700 bp | 450 bp |
| Typical Throughput | 700 Mb | 450 Mb |
| Reads per Run | ~1,000,000 shotgun | ~1,000,000 shotgun or 700,000 amplicon |
| Run Time | 23 hours | 10 hours |
| Sample Input | gDNA or cDNA | gDNA, cDNA, or amplicons (PCR products) |
| Multiplexing | Multiplex Identifiers (MIDs): 132 Gaskets: 2, 4, 8 regions | Multiplex Identifiers (MIDs): 132 Gaskets: 2, 4, 8 regions |
Under optimal conditions, ~630-910 million bases of an average read length of ~700 bases and can be obtained with FLXPLUS Chemistry and
about ~400-600 million bases of an average read length of ~400 bases and can be obtained with Titanium Chemistry.
Applications
- De Novo Sequencing: Whole Genomes of Humans, Plants, Yeasts, Fungi, Bacteria, Viruses, BACS etc.
- Resequencing: SNPs, InDels, Somatic Mutations
- Transcriptome Analysis: cDNA, small non coding RNA, microRNA
- Epigenetic Changes: DNA Methylation Patterns
- Metagenomics: Analysis of Environmental DNA
- Paleogenomics : Ancient DNA
DNA Requirements
2-4µg of double stranded DNA in more than 10µl volume is required.Ideally DNA size should be larger than 2kb. If your DNA is 500bp long or less, then about 3µg in more than 10µl volume is necessary. The DNA should be suspended in H2O or 1X TE.
Provide the DNA at a concentration > 300ng/µl and A260/280 ratio of approximately 1.8.
An image of DNA sample(s) run on a 1-2% agarose gel is requested to assess quality of incoming DNA. We will further assess the quality using Bioanalyzer and Nanodrop.
De novo sequencing requires 20X average genome coverage. Resequencing and mapping projects require 15X average coverage.
Application notes/Protocols
- Amplicon Sequencing
- Metagenomics
- SAGE
- Methylation
- De Novo Bacteria Sequencing
- Paired End Reads
- Cloning Small RNAs for Sequencing
- RNA Virus Sequencing
High Throughput Sequencing Services Contact Info
- The Sequencing and Genotyping Core is located at
- 5309 Gonda Center on the UCLA campus.
- 695 Charles Young Drive South
- Los Angeles, CA 90095
- "Gonda Neurosci Genet Res Ctr" on the UCLA Campus Map
- Open 9AM to 5PM Mon-Fri, excluding university holidays
- For more information about sequencing at the Core, call 310-267-2461 or contact:
- Uma Dandekar, GenoSeq Core Assistant Director
- Email: uma@ucla.edu
- Phone: 310-267-2461
- Fax: 310-794-5446