Libraries prepared with ThruPLEX Plasma-seq provide added sensitivity to better understand genetic variations found within the plasma samples. Plasma, along with other liquid biopsies, has gained significant interest in the research community as a valuable and easily available sample type. These communities include translational research in oncology and developmental research involving the fetal fraction derived from maternal plasma.
Highest Diversity and Fewest Unmapped Reads from Cell-Free DNA
ThruPLEX Plasma-seq generated quality libraries with high diversity and a low number of duplicates and unmapped reads. Cell-free DNA was extracted from 3 plasma samples and libraries were prepared at the amounts indicated as measured by Qubit®. The amount of mononucleosomal DNA in each sample, as measured by the Bioanalyzer®, corresponded to 0.09 ng, 0.62 ng, and 15.44 ng. Pooled libraries were sequenced on an Illumina NextSeq® 500 as a paired-end run with 17M to 25M reads per library. Duplication rates were calculated after down-sampling the data to 17 M reads per library.
Reproducible, Unbiased GC Coverage
ThruPLEX Plasma-seq provided the most reproducible and unbiased GC coverage across the human genome. ThruPLEX libraries showed minimal variability across 9 individual plasma samples tested. Libraries were prepared from cell-free DNA isolated from 1 mL of plasma samples and sequenced on an Illumina NextSeq 500. Four separate plasma samples were used to construct the NEBNext® Ultra™ libraries.
Outstanding Target Enrichment Performance
ThruPLEX Plasma-seq libraries were captured at high efficiency and generated data with deep coverage of the kinome for mutation detection. Libraries were prepared from 3 plasma samples at input amounts of 5 ng, 6.5 ng, and 10 ng in triplicate, and targeted sequencing was carried out on an Illumina MiSeq® using samples enriched with the ClearSeq® Human DNA Kinome Panel for SureSelectXT2. On average, 5 M reads were generated per library. Selected bases were successfully captured bases that were in or within 250 bp of the baits.
- Designed for cell-free DNA - Newly formulated repair and ligation reagents
- High Performance NGS Libraries - High library diversity at low input amount with broad and reproducible GC coverage
- Adaptable Sample Input Amount - <1 to 30 ng of cell-free DNA
- Fast and Simple Workflow - 3 steps in a single tube or well in 2 hours with no purification or sample transfer steps
- Demonstrated method on Biomek FXP Automated Workstation