We performed whole-genome sequencing on extracted DNA samples following the Illumina DNA Prep manufacturer protocol (Illumina, United States). We constructed and pooled libraries using the Illumina DNA Prep Kit. We measured library concentrations on a Qubit4 Flourometer using the Qubit High Sensitivity 1x dsDNA kit (ThermoFisher Scientific, United States), while we measured the average library size on an Agilent TapeStation 4150 using the Agilent High Sensitivity D1000 ScreenTape kit (Agilent Technologies, United States). We conducted genomic sequencing on an Illumina MiniSeq instrument (Illumina, United States).
Qubit4 flourometer
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Qubit4 Fluorometer is a compact and sensitive instrument designed for the accurate quantification of DNA, RNA, and protein samples. It utilizes fluorescence-based detection to provide precise and reliable measurements of sample concentrations.
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Lab products found in correlation
Agilent tapestation 4150, by Agilent Technologies (1 mentions)
Dna prep kit, by Illumina (1 mentions)
Miniseq, by Illumina (1 mentions)
Agilent high sensitivity d1000 screentape kit, by Agilent Technologies (1 mentions)
Dreamtaq dna polymerase, by Thermo Fisher Scientific (1 mentions)
Qiaquick pcr purification kit, by Qiagen (1 mentions)
Novaseq platform, by Illumina (1 mentions)
Qubit dsdna br assay, by Thermo Fisher Scientific (1 mentions)
2 protocols using qubit4 flourometer
1
Whole-Genome Sequencing of DNA Samples
2
Detecting Mosaic Genetic Variants
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The putative mosaic variants were targeted by PCR primers designed using the Primer3Plus tool. Candidate mosaic parental samples were amplified by PCR using Dreamtaq DNA polymerase (Thermofisher Scientific, Waltham, MA, USA). PCR products for each parental sample were purified using the QIAquick PCR Purification Kit (Qiagen) following the manufacturer’s protocol. The concentration of the PCR products was quantified using the Qubit dsDNA BR Assay (ThermoFisher Scientific) by the Qubit 4 Flourometer (ThermoFisher Scientific). Purified PCR products of 150–280 bp in length were sequenced using the Illumina Novaseq platform (Illumina, San Diego, CA, USA) with 150-bp paired end reads at Cloud Health Genomics (Shanghai, China). Data were analyzed using BWA-mem, Integrative Genomics Viewer (IGV), and GATK HaplotypeCaller for putative mosaic indels. Custom scripts were written in R statistical programming language.
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