Illumina Genome Analyzer is a new generation high-throughput sequencing platform based on formation of clonal molecule clusters and the principle of SBS (synthesis-based-sequencing). It can sequence millions of fragments using proprietary reversible terminator-based sequencing chemistry per reaction.

This sequencing technology relies on the attachment of randomly fragmented genomic DNA to a planar, optically transparent surface. Attached DNA fragments are extended and bridge amplified to create a sequencing flow cell with hundreds of millions of clusters, each containing ~1,000 copies of the same template. These templates are sequenced using a robust four-color DNA SBS (sequencing-by-synthesis) technology that employs reversible terminators with removable fluorescent dyes. This novel approach ensures high accuracy and true base-by-base sequencing, eliminating sequence-context specific errors and enabling sequencing through homopolymers and repetitive sequences.

This technology overcomes the shortcomings of the traditional sequencing techniques as high cost in labor and materials, and the need of cloning, transformation and plasmid preparation. Alternative sample preparation methods allow a range of applications of this sequencing technology including gene expression, small RNA discovery, meta-genomics, DNA methylation and protein-nucleic acid interactions.

Experimental workflow:

1.DNA library construction

--Genomic DNA fragmentation

--End-repair of fragments

--Adapter ligated to 3’ and 5’ ends

2.Cluster generation on Cluster Station.

--Flow cell and pre-filled reagents placed into Cluster Station

--DNA samples applied to flow cell

                                                     --Complete walk-away automation

3.SBS (synthesis-based-sequencing) on Genome Analyzer.

--Flow cell and pre-filled reagents placed into Genome Analyzer

--Complete walk-away automation, including support for longer reads

--Image processing, real-time analysis (RTA), base calling

4.Paired-End module

--Add-on module for automated reagent delivery

--In situ regeneration of template and preparation for second read

5.Post-Run bioinformatics analysis

--Alignment and assembly of sequences

--Various analysis (e.g., SNP, methylation, expression profiling)

--Summary report

Features:

 • High-throughput. The Illumina Genome Analyzer can generate data over 3G per run.

 • High accuracy.  Accuracy is over 98.5% and good for tandem repeat sequences.

 • Low cost. Lower than 1% of the traditional capillary sequencing technique.

 • Long read. DNA sequence read up to 200 bp.

 • Paired-end module. Sequencing from reverse directions with insert ranging in size from 200 bp up to 10 kbp.

 • Maximum production. Each flowcell has 8 channels. Each channel can run one or a mixed sample.