Reduced PCR cycling time amplification using AmpFℓSTR ® Identifiler ® kit

To reduce the DNA profiling time, DNA samples were amplified using AmpFlSTR® Identifiler® 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

Reduced volume PCR amplification using AmpFℓSTR ® Identifiler ® kit

This study was aimed to demonstrate that the AmpFlSTR® Identifiler® kit will reliably amplify DNA in a reaction volume of 10ml. For comparison, the DNA samples were also amplified at the currect in house PCR reaction volume (12.5ml). Even though the results showed significant difference in the peak height intensities, the quality of the profiles produced in the reaction volume of 10ml was similar to those produced in the reaction volume of 12.5ml. 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

100,000 Genomes Pilot on Rare-Disease Diagnosis in Health Care - Preliminary Report.

BACKGROUND: The U.K. 100,000 Genomes Project is in the process of investigating the role of genome sequencing in patients with undiagnosed rare diseases after usual care and the alignment of this research with health care implementation in the U.K. National Health Service. Other parts of this project focus on patients with cancer and infection. METHODS: We conducted a pilot study involving 4660 participants from 2183 families, among whom 161 disorders covering a broad spectrum of rare diseases were present. We collected data on clinical features with the use of Human Phenotype Ontology terms, undertook genome sequencing, applied automated variant prioritization on the basis of applied virtual gene panels and phenotypes, and identified novel pathogenic variants through research analysis. RESULTS: Diagnostic yields varied among family structures and were highest in family trios (both parents and a proband) and families with larger pedigrees. Diagnostic yields were much higher for disorders likely to have a monogenic cause (35%) than for disorders likely to have a complex cause (11%). Diagnostic yields for intellectual disability, hearing disorders, and vision disorders ranged from 40 to 55%. We made genetic diagnoses in 25% of the probands. A total of 14% of the diagnoses were made by means of the combination of research and automated approaches, which was critical for cases in which we found etiologic noncoding, structural, and mitochondrial genome variants and coding variants poorly covered by exome sequencing. Cohortwide burden testing across 57,000 genomes enabled the discovery of three new disease genes and 19 new associations. Of the genetic diagnoses that we made, 25% had immediate ramifications for clinical decision making for the patients or their relatives. CONCLUSIONS: Our pilot study of genome sequencing in a national health care system showed an increase in diagnostic yield across a range of rare diseases. (Funded by the National Institute for Health Research and others.)