Ultima UG100 Whole Genome  single-end 300bp Sequencing of the GIAB HG008-N-D normal duodenal tissue
Sequencing was performed on the HG008-T pancreatic ductal adenocarcinoma tumor cell line with matched normal duodenal tissue (HG008-N-D), using similar methods and library preparation protocols tailored for both tumor and normal samples, as described below. The data presented here corresponds to the normal duodenal tissue (HG008-N-D).Genomic DNA (gDNA) from non-viable normal tissue was isolated using the Qiagen Puregene Tissue Kit (PN: 158667).  Approximately 2 ml of Qiagen kit cell lysis buffer with 30 ul of Proteinase K was added to 10-15 mg of tissue. A Dounce Homogenizer was used to disrupt the tissue before leaving it overnight at 55C. This was followed by addition of RNase A and another incubation at 37C for 5 minutes followed by cooling on ice. Genomic DNA was precipitated, washed and hydrated in TE buffer (1 mM EDTA, 10 mM TrisCl pH 7.5) per the Qiagen Puregene protocol.. Library preparation for each sample, started with 500 ng of isolated genomic DNA.  PCR-free library preparation was performed using the NEBNext Ultra II FS DNA PCR-free Library Prep Kit (PNs: E7805, E6177) following Ultima Genomics NEB PCR-Free WGS Library Preparation Protocol (Publication #: P00014 Rev. D).  Ultima Genomics PCR-free adapters, xGen PCR-Free Adapters for Ultima P1, 96 rxn (PN: 10016841) for UG 100 sequencer compatibility were used.Sequencing was performed on the UG 100 Sequencer (software version : APL 5.1.0.19) using iV27 Chemstry. Single end, 300 bp reads were generated for both samples using the UG_116cycles_Baseline_1.5.3.2  sequencing recipe and flow order [T,G,C,A] on a single wafer. A human genome control sample HG002 (Coriell Nucleic acid ID: NA24385) was spiked into each run in order to ensure quality of the samples being sequenced.