Library construction for next-generation sequencing: Overviews and challenges

By S. Head, H. Komori, Sarah A. LaMere, T. Whisenant, F. van Nieuwerburgh, D. Salomon, Phillip T. Ordoukhanian. BioTechniques, 2014

620 citations recorded by the source corpus.

High-throughput sequencing, also known as next-generation sequencing (NGS), has revolutionized genomic research. In recent years, NGS technology has steadily improved, with costs dropping and the number and range of sequencing applications increasing exponentially. Here, we examine the critical role of sequencing library quality and consider important challenges when preparing NGS libraries from DNA and RNA sources. Factors such as the quantity and physical characteristics of the RNA or DNA source material as well as the desired application (i.e., genome sequencing, targeted sequencing, RNA-seq, ChIP-seq, RIP-seq, and methylation) are addressed in the context of preparing high quality sequencing libraries. In addition, the current methods for preparing NGS libraries from single cells are also discussed.

Subfields: Molecular Biology

View Paper: https://www.semanticscholar.org/paper/03c6fe49da90735c1e43023811d29d5fa6cd5820

How researchers see this paper

The researchers whose trusted networks rank it highest, strongest advocate first. Every researcher gets a different order.

In Molecular Biology

Citations in the corpus

Updated Molecular Testing Guideline for the Selection of Lung Cancer Patients for Treatment With Targeted Tyrosine Kinase Inhibitors: Guideline From the College of American Pathologists, the International Association for the Study of Lung Cancer, and the Association for Molecular Pathology (2018, background): “of malignant cells, and, although typically slower than 1 single-gene assay, can often be performed more rapidly than sequential multiple single-gene assays.(175,176) A”
Adapterama I: universal stubs and primers for 384 unique dual-indexed or 147,456 combinatorially-indexed Illumina libraries (iTru & iNext) (2019, methodology): “A variety of library preparation methods have also been described by research groups that reduce per-sample costs relative to most commercial kits (e.g., Meyer & Kircher, 2010 [MK-2010]; Fisher et al., 2011 [F-2011]; see Head et al., 2014 for others).”
Single‐tube library preparation for degraded DNA (2017, background): “However, these preparations often lead to biases in GCcontent, fragment length, or complexity (Aird et al. 2011; Dabney & Meyer, 2012; Seguin-Orlando et al. 2013; Head et al. 2014; Van Dijk et al. 2014).”
DNA Sequencing as a Tool to Monitor Marine Ecological Status (2017, methodology): “In general, DNA extracted from a sample is fragmented, a DNA library is prepared (Head et al., 2014) using a limited number of PCR cycles or via PCR-free methods (Mendoza et al.”