Dsdna quantifluor system kit

Sera-Mag™ SpeedBeads was prepared according to Faircloth et al. [27 ] and used to clean the PCR products using a Zephyr NGS Workstation (Caliper Lifesciences, Perkin-Elmer) and quantified using a Promega dsDNA Quantifluor System Kit (Ref: E2670, 00002484139) on an Enspire Workstation (Perkin-Elmer). The 5 different PCR products were pooled at approximately 0.4 nM to ensure even coverage during sequencing using Quantifluor dsDNA System (Promega, Madison, WI, USA). The products were normalized to 2 ng/µL using 10 mM Tris-HCl (pH 8.0). Final dilution to 0.2 ng/µL with 10 mM Tris-HCl (pH 8.0) was conducted in preparation for library preparation and final accuracy checks using the Illumina Nextera^XT DNA.

PCR products were cleaned using Sera-Mag™ SpeedBeads (Merck KGaA, Darmstadt, Germany) in a Zephyr NGS Workstation (Caliper Lifesciences, Perkin-Elmer; Waltham, MA, USA) and quantified using a Promega dsDNA Quantifluor System Kit (Ref: E2670, 00002484139; Madison, WI, USA) on an Enspire Workstation (Perkin-Elmer; Waltham, MA, USA). The PCR products of all fragments were pooled and normalized to 0.2 ng/µL with 10 mM Tris-HCl (pH 8.0) for library preparation, and final accuracy checks of concentration were performed using the Illumina Nextera XT DNA Library Prep Kit (Illumina, San Diego, CA, USA).