Whole exome sequencing was performed on DNA extracted from whole blood using previously described method with the following alterations: the Agilent SureSelectXT Human All Exon V4 kit (Agilent Technologies) was used29 (link), and sequencing was performed on the HiSeq2500 instrument (Illumina) with 100bp paired-end sequencing. Sequence was aligned to the human reference genome (Hg19) using Burrow-Wheeler Aligner (0.5.11)30 (link); variants were called with Samtools31 (link) and Genome Analysis Toolkit (v.1.6)30 (link) and annotated with Annovar31 (link). Variants with a quality score <10 were excluded from analysis. Variant filtering and segregation was performed with in house developed software (https://github.com/brendanofallon/VarViewer ). All genes previously associated with congenital lymphedema (CCBE1, FOXC2, FLT4, KIF11, GATA2, GJC2, SOX18, FAT4 and PTPN14) were specifically analyzed and no rare or pathogenic variants were found in any family member despite full sequencing coverage. The reported PIEZO1 variants were confirmed by Sanger sequencing using standard methods. Primer sequences are available upon request.
Agilent sureselectxt human all exon v4 kit
Manufactured by Agilent Technologies
The Agilent SureSelectXT Human All Exon V4 kit is a targeted enrichment solution for next-generation sequencing (NGS) applications. The kit is designed to capture the complete set of human protein-coding exons, providing a comprehensive coverage of the human exome. The kit includes pre-designed RNA baits that target known exonic regions, enabling efficient and specific enrichment of the human exome from DNA samples.
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Lab products found in correlation
2 protocols using agilent sureselectxt human all exon v4 kit
1
Whole Exome Sequencing for Congenital Lymphedema
2
Whole-Exome Sequencing for Congenital Lymphoedema
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Whole-exome sequencing was performed on DNA extracted from whole blood using a previously described method with the following alterations: the Agilent SureSelectXT Human All Exon V4 kit (Agilent Technologies) was used27 (link), and sequencing was performed on the HiSeq2500 instrument (Illumina) with 100-bp paired-end sequencing. Sequence was aligned to the human reference genome (Hg19) using Burrow–Wheeler Aligner (0.5.11)28 (link); variants were called with Samtools29 (link) and Genome Analysis Toolkit (v.1.6)30 (link) and annotated with Annovar31 (link). Variants with a quality score <10 were excluded from analysis. Variant filtering and segregation was performed with in-house developed software (https://github.com/brendanofallon/VarViewer ). All genes previously associated with congenital lymphoedema (CCBE1, FOXC2, FLT4, KIF11, GATA2, GJC2, SOX18, FAT4 and PTPN14) were specifically analysed and no rare or pathogenic variants were found in any family member despite full sequencing coverage. The reported PIEZO1 variants were confirmed by Sanger sequencing using standard methods. Primer sequences are available upon request.
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