Exome Sequencing of Affected Families

Genomic DNA was extracted from whole blood from the affected children and their parents. Exome sequencing at GeneDx was performed on exon targets captured using the Agilent SureSelect Human All Exon V4 (50 Mb) or the Clinical Research Exome kit (Agilent Technologies) according to the manufacturer's instructions. Libraries were sequenced using the Illumina HiSeq 2000 or 2500 sequencing system with 100-bp paired-end reads (Illumina). Whole-exome sequence data for all sequenced family members was analyzed using GeneDx's XomeAnalyzer (a variant annotation, filtering, and viewing interface for exome sequencing data) as described previously (Tanaka et al. 2017 (link)). The identified sequence variant of interest was confirmed in the proband, affected siblings, and both parents by conventional di-deoxy DNA sequence analysis using an ABI3730 (Life Technologies).

Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link: Access Free Full Text.

Tanaka A.J., Okumoto K., Tamura S., Abe Y., Hirsch Y., Deng L., Ekstein J., Chung W.K, & Fujiki Y. (2019). A newly identified mutation in the PEX26 gene is associated with a milder form of Zellweger spectrum disorder. Cold Spring Harbor Molecular Case Studies, 5(1), a003483.

Publication 2019

SureSelect Human All Exon V4 (50 Mb) Clinical Research Exome kit HiSeq 2000 or 2500 sequencing system with 100-bp paired-end reads ABI3730

Blood Bp 100 Children Deoxy Dna sequence analysis Exome Exon Family members Genomic Human Parents Siblings

Corresponding Organization :

Other organizations : Columbia University Irving Medical Center, Kyushu University

Top 5 similar protocols

Variable analysis

independent variables

Exome sequencing at GeneDx performed on exon targets captured using the Agilent SureSelect Human All Exon V4 (50 Mb) or the Clinical Research Exome kit (Agilent Technologies)

dependent variables

Whole-exome sequence data for all sequenced family members analyzed using GeneDx's XomeAnalyzer
Identified sequence variant of interest confirmed in the proband, affected siblings, and both parents by conventional di-deoxy DNA sequence analysis using an ABI3730 (Life Technologies)

control variables

Genomic DNA extracted from whole blood from the affected children and their parents

positive controls

None specified

negative controls

None specified

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!