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GS FLX+ System  : High Throughput Sequencer

454 Sequencing Page

GS FLX

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


Publications:



High Throughput Sequencing Services Contact Info

The Sequencing and Genotyping Core is located at
36-125 CHS
UCLA Center for Health Sciences Building
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
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