Protocol detail

Exome Sequencing from Blood Samples

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DNA was extracted from blood cells using a FlexiGene DNA kit (Qiagen, Valencia, USA). Exome sequences were collected according to previously described procedures [15 (link)]. Briefly, genomic DNA was fragmented using a Covaris II system (Covaris, Woburn, MA, USA). Exon templates were isolated using the TruSeq Exome Enrichment Kit (Illumina, San Diego, CA, USA) and exome sequences were collected in a HiSeq2500 sequencer (paired-end 2×150) at 100x coverage. The total variants called from the exome data from this study have been deposited in DRYAD Digital Repository with accession ID: doi:10.5061/dryad.p236p.

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Kim Y.C., Soliman A.S., Cui J., Ramadan M., Hablas A., Abouelhoda M., Hussien N., Ahmed O., Zekri A.R., Seifeldin I.A, & Wang S.M. (2017). Unique Features of Germline Variation in Five Egyptian Familial Breast Cancer Families Revealed by Exome Sequencing. PLoS ONE, 12(1), e0167581.

Publication 2017

FlexiGene DNA kit Covaris II system TruSeq Exome Enrichment Kit HiSeq2500 sequencer

Blood cells Exome Exon Genomic

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Variable analysis

independent variables

Extraction of DNA from blood cells using a FlexiGene DNA kit
Fragmentation of genomic DNA using a Covaris II system
Isolation of exon templates using the TruSeq Exome Enrichment Kit
Sequencing of exome sequences in a HiSeq2500 sequencer (paired-end 2×150) at 100x coverage

dependent variables

Exome sequences collected

control variables

Procedures previously described in reference 15

controls

No positive or negative controls were explicitly mentioned in the provided information.

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