Kapa sybr fast qpcr with

Isolated nucleic acid was quantified with Qubit 2.0 DNA HS Assay (Thermo Fisher) and the quality was assessed by Tapestation Genomic DNA Assay (Agilent Technologies, CA, USA). Library preparation was performed using the NexteraXT library kit (Illumina, CA, USA) according to the manufacturer’s recommendations. The final library quantity was measured by KAPA SYBR^® FAST qPCR with QuantStudio^® 5 System (Applied Biosystems, CA, USA), and library quality was evaluated by TapeStation HSD1000 ScreenTape (Agilent Technologies). Illumina^® 8-nt dual indices were used. Equimolar pooling of libraries in the same run was performed based on QC values and sequenced on an Illumina^® NovaSeq with a read length configuration of 150 paired-end.

Library preparation and sequencing were performed by Admera Health (South Plainfield, NJ, USA). Isolated genomic DNA was quantified with Qubit 2.0 DNA HS Assay (ThermoFisher, Waltham, MA, USA) and DNA quality was evaluated by Genomic DNA ScreenTape analysis (Agilent Technologies, Santa Clara, CA, USA). Library preparation was performed using Nextera XT library kit (Illumina, San Diego, CA, USA) following the manufacturer’s recommendations while barcoding was performed using Illumina 8-nt dual-indices. The final library was evaluated by Qubit 2.0 DNA HS Assay and the quality of the library generated was examined by TapeStation HSD1000 ScreenTape (Agilent Technologies). Final library quantity was measured by KAPA SYBR FAST qPCR with QuantStudio 5 System (Applied Biosystems, Waltham, MA, USA). Equimolar pooling of libraries was performed based on qPCR QC values before sequencing on an Illumina HiSeq with a read length configuration of 2 × 150 paired end reads, obtaining 1 M PE reads per sample.