Platforms
AGRF's Next-Generation Sequencing service has the capability and experience to process sequencing projects that utilise many applications. AGRF utilises two platforms for it's next-generation sequencing projects: the Roche GS FLX and the illumina GA II. The GS FLX generates longer read lengths for applications including de novo genome sequencing and re-sequencing. Applications that use short reads, but require greater sequencing depth, include ChIP-Seq which can be performed on the illumina GA II.
Next-Generation Sequencing Service Specifications
|
Parameter |
Roche GS FLX |
illumina GA II |
|
Sequencing Output
|
300 - 500 Mb per full plate run
|
· >1.5 Gb per 1 lane run (single read) · >3.0 Gb per 1 lane run (paired end) |
|
Number of Reads
|
750 000 per full plate run |
10 000 000 per 1 lane run |
|
Average Read length
|
250 - 400 bp
|
· Up to 76 bp · Up to 76 bp x 2 (paired end) |
|
Raw Read Accuracy |
>99.5% |
>98.5% |
|
Throughput
|
· Up to 16 libraries per plate · Barcoding options available |
· 7 channels per flow cell · Up to 12 samples per channel |
|
AGRF Input Requirements
|
· 1 - 2 μg genomic DNA · >100 ng PCR-generated template |
· 0.1 - 5.0 μg (single and paired end reads) · 10 - 20 μg (Mate Pair Reads) · 1 - 10 μg (total RNA) |
Roche GS FLX
AGRF utilises the Roche GS FLX sequencing platform, providing unparalleled read capacity with state-of-the-art sequencing technology. Many applications have been developed for this instrument, including the large scale analysis of bacterial/yeast genomes, BAC pools, transcriptomes, microRNA libraries and ultra deep Amplicon sequencing. In addition, the new application of paired-end sequencing enables contig alignment, greatly reducing the time for bacterial/yeast genome finishing. The GS FLX is capable of producing up to 1,250,000 reads per run with an average read length between 200 and 400 bp or up to 500 Mb total sequence at 99.5% raw base accuracy. The AGRF's Bioinformatics team utilises leading sequence analysis software enabling high quality base calling and flexibility to accommodate a variety of applications.
illumina GA II
AGRF utilises the illumina GA II sequencing platform, which uses a massively parallel sequencing-by-synthesis approach to generate billions of bases of high-quality DNA sequence per run. Many applications have been developed for this instrument, including RNA Sequencing, and ChIP-Seq. The GA II is capable of producing up to 20,000, 000 reads per 1 lane run with an average read length of up to 76 bp or >1.5Gb total sequence at 98.5% raw base accuracy. The AGRF's Bioinformatics team utilises leading sequence analysis software enabling high quality base calling and flexibility to accommodate a variety of applications.
