Technical Specifications

Work Flow

• Library Preparation: Each application requires specific sample preparation for the sequencing and sample library is prepared according to the recommended Illumina protocol for each application.
• Library Validation: For routine library validation, Agilent 2100 bioanalyzer is used to ascertain the library fragment size, yield and concentration. We offer additional option for library quality control using cloning and capillary sequencing.
• Cluster Generation: During cluster generation, the library hybridises to the flow cell and series of bridge amplification occurs, resulting in the generation of approximately 1000 copies for each template. We expect to see up to 25 million high quality clusters formed on each lane of the flow cell.
• Sequencing by Synthesis: Each nucleotide is designed to have a reversible termination property, which allows each cycle of the sequencing reaction to occur simultaneously in the presence of all four nucleotides. Read lengths from 38bp to 100bp are available in both single or paired end reads.

Sample Requirements

The input material that we require exceeds the amounts required for sample preparation. The additional amount is necessary to perform initial QC of the samples. Please contact us if your samples do not meet the requirements.

We will perform an initial QC of all sample material as follows:

If the samples received do not meet our QC requirements, we will contact you to discuss how to proceed.

Deliverables

For a good sequencing run, up to 25 million reads may now be reliably detected (pass filter). In addition, the latest innovations in image analysis and cluster detection software mean that more and more distinct clusters may be identified per tile, thus leading to even greater yields.

Once all analysis has been completed, all data is returned to the customer on external hard drive. This will include:

• All intermediate files generated during analysis. Retention of this data by the customer allows further experiments to be carried out in the future, without needing to re-sequence samples
• Final (filtered) output data, in FASTA or FASTQ format. FASTQ is a modification of the popular FASTA format, with quality scores assigned to each base-call.
• Data from all bioinformatics analysis that have been performed. File formats will depend on the particular type of analysis.
• Lab QC report detailing the integrity of the samples that were used for sequencing.
• Bioinformatics pipeline report, explaining quality control metrics used to assess the quality of the data, and details of all bioinformatics analyses that have been performed, explaining the methods used and results obtained.