The Illumina HiSeq 2500 sequencing services provided by the Genome Sequencing Facility (GSF) allows for both the standard High Output 8 lane flow cell run and the 2 lane flow cell Rapid Run. Both services are supported by Paired-End (PE) or Single Read (SR) sequencing using the Illumina TruSeq SBS - HS and TruSeq Rapid SBS - HS chemistry. The High Output runs are available indexed or non-indexed as 50 or 100 cycle PE or SR. The Rapid Run is available indexed or non-indexed as 50, 100 or 150 cycle PE or SR. Flow cell clustering is performed using the cBOT cluster generation station. The following information is a comparison of expected output and data quality for both High Output and Rapid Read runs.
Rapid (2 lane) High Output (8 lane)
Cluster Density 750-900 k/mm2 750-850 k/mm2
Clusters/Lane (PF) 130 - 150M 187 - 210M
Flow Cell Output (2x50bp) 25 - 30Gb 150Gb
Flow Cell Output (2x100bp) 50 - 60Gb 300Gb
Flow Cell Output (2x150bp) 75 - 90Gb not supported
Bases >Q30 (2x50bp) 85% 85%
Bases >Q30 (2x100bp) 80% 80%
Bases >Q30 (2x150bp) 75% not supported
Run times between the Rapid Read and High Output are significantly different. The following provides a comparison of run times.
Run Format Rapid Read High Output
50 ~ 8 hours ~ 2 days
2 x 50 ~ 16 hours ~ 5 days
2 x 100 ~ 27 hours ~ 10 days
2 x 150 ~ 40 hours not supported
Sequencing library construction is also available through the GSF using the Illumina TruSeq Sample preparation kits. The following library services are offered:
TruSeq mRNA - Generate mRNA libraries directly from total RNA.
TruSeq Stranded mRNA - Generate mRNA libraries directly from total RNA that provide strand origin for sequenced mRNA transcripts.
TruSeq DNA - Generate Libraries from Genomic DNA.
TruSeq ChIP-Seq DNA - Selectively sequence DNA sequences bound by target proteins.
TruSeq Small RNA - Generate small RNA libraries directly from total RNA.
TruSeq Exome - Ideal for scalable exome sequencing studies.
For other library constructs please inquire.
Mar 07, 2014