Services

Cluster generation and sequencing

After QC, prepared DNA libraries can be submitted to the Facility for cluster generation and sequencing.
The Facility has two Illumina Genome Analyzer-IIx units; each is :

1. capable of producing reads as long as 150-bp;
2. connected to a module for Paired-End (PE) runs.

Before a sample can be run on the Genome Analyzer, analysis and read mapping must be discussed with Babraham Bioinformatics.

Quality control and quantitation

Quality control (QC) before Illumina sequencing is crucial to maximizing the amount of data obtained from a run. QC to determine the size distribution of fragments in the DNA library is done using an Agilent Bioanalyzer 2100, a ‘lab-on-a-chip’ microfluidics system for sizing and quantifying DNA.

The Bioanalyzer High Sensitivity Assay:

1. detects a minimum of 5 pg/µL of DNA (assuming a uniform fragment size);
2. sizes fragments in a range of 50 - 10,000 bp;
3. takes 45 minutes to run.

Wells on a Bioanalyzer chip may be booked. Samples that pass QC are then quantitated using a Taqman real-time PCR assay, as described by Quail et al. 2008 (see References).

Sample preparation kits and protocols

Illumina reagents and protocols can be used to prepare libraries for Next Generation Sequencing.
Protocols for sample preparation are available on request