Background The UK 100,000 Genomes Project is in the process of investigating the role of genome sequencing of patients with undiagnosed rare disease following usual care, and the alignment of research with healthcare implementation in the UK’s national health service. (Other parts of this Project focus on patients with cancer and infection.) Methods We enrolled participants, collected clinical features with human phenotype ontology terms, undertook genome sequencing and applied automated variant prioritization based on virtual gene panels (PanelApp) and phenotypes (Exomiser), alongside identification of novel pathogenic variants through research analysis. We report results on a pilot study of 4660 participants from 2183 families with 161 disorders covering a broad spectrum of rare disease. Results Diagnostic yields varied by family structure and were highest in trios and larger pedigrees. Likely monogenic disorders had much higher diagnostic yields (35%) with intellectual disability, hearing and vision disorders, achieving yields between 40 and 55%. Those with more complex etiologies had an overall 25% yield. Combining research and automated approaches was critical to 14% of diagnoses in which we found etiologic non-coding, structural and mitochondrial genome variants and coding variants poorly covered by exome sequencing. Cohort-wide burden testing across 57,000 genomes enabled discovery of 3 new disease genes and 19 novel associations. Of the genetic diagnoses that we made, 24% had immediate ramifications for the clinical decision-making for the patient or their relatives. Conclusion Our pilot study of genome sequencing in a national health care system demonstrates diagnostic uplift across a range of rare diseases. (Funded by National Institute for Health Research and others)

This study was aimed to demonstrate that the AmpF'STR 1 Identifiler 1 kit will reliably amplify DNA in a reaction volume of 10 ml. For comparison, the DNA samples were also amplified at the currect in-house PCR reaction volume (12.5 ml). Even though the results showed significant difference in the peak height intensities, the quality of the profiles produced in the reaction volume of 10 ml was similar to those produced in the reaction volume of 12.5 ml. The application of this reduced volume PCR amplification would represent an additional 20% cost saving on the reagents without compromising the quality of the profiles obtained.2015 Elsevier Ireland Ltd. All rights reserved.

A B S T R A C TTo reduce the DNA profiling time, DNA samples were amplified using AmpF'STR 1 Identifiler 1 kit at reduced PCR cycling time amplification method without addition of any enhancement buffers or enzymes. The results showed no significant difference in the quality of the profiles produced compared to those produced in the manufacturer recommended condition. This study contributed in saving almost 25% of the time needed for PCR amplification without compromising the quality of the profiles.