Validation Data on VERSA Gene Workstation
- Table 1 – Purified amplified NGS library concentrations for positive and negative controls for both manual and automated preparations.
- Table 2 – Correlation of sequencing data obtained by manual and automated NGS library preparations.
What are the advantages of Versa 10 and 1100 automated Illumina NGS Library construction?
- More efficient bead mixing limits bead clumping and wastage as a result of beads attaching to the pipette tip and reservoir.
- Bead distribution is no longer time-sensitive because the beads are kept in constant suspension versus the risk of bead settling in a reservoir.
- Minimizes dead volume as magnetic beads are dispensed from the source tube and not a reservoir.
- Validating Magnetic Bead Vortex In every NGS library preparation protocol, magnetic beads play an essential role and thus have a direct impact on the yield, concentration, consistency and quality of NGS library. Therefore, uniform bead distribution and accurate dispensing is critical.
- Purified amplified NGS library concentrations for positive and negative controls for both manual and automated procedures within the acceptable quality control limits as outlined by the end-user.
- Correlation (r ) of sequencing data obtained by manual and automated NGS library preparation procedures suggesting that automated procedures produce high quality sequencing data (Table 2).
- Sequencing quality and reproducible can be attributed to the uniform suspension and accurate dispensing on magnetic beads by the VERSA Gene magnetic bead vortex and automated liquid handling modules.
Library preparation on the VERSA 10 or 1100 Gene Workstation
- NGS data from Ion AmpliSeq Cancer Hotspot Panel
- The checker board experiments concluded that VERSA automated liquid handling system showed no evidence of cross-contamination, by either no library in the NTC wells, or no variants called on negative samples after sequencing the CHP2 assay.
- High reproducibility observed in both library yields and variants called across all technical replicates of the quality control materials.
- All patient DNA samples yielded good quality libraries, including those difficult samples that had previously failed in the manual library preparation method.
- The variants called with highly correlated (Pearson’s r >0.990) frequencies to those obtained with the manual method as shown on the right by representative curve on the 36 variants frequency identified in the positive control sample by both library preparation methods.
Scientific NGS / Aurora References:
1. Gill et al. (2015). Validation of a Magnetic Bead Mixer on an Automated Next-Generation Sequencing Library Preparation System. American Laboratory.
2. Dumur et al. (2015, November). Validation of an Automated Method for Library Preparation for a Next Generation Sequencing-Based Assay for Oncology. Poster presented at American Molecular Pathology, Austin, TX.
Data on validation of magnetic bead vortex and its contribution to NGS library preparation alone with NGS data on Cancer Hotspot Panel