ArticleNucleic acids research2017
Single-stranded DNA library preparation from highly degraded DNA using T4 DNA ligase.
Article in Nucleic acids research, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 136 papers.
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136 citing papers in PubMed, 245 citations in OpenAlex.
- Comparing the Performance of Double-Stranded and Single-Stranded DNA Libraries for Ancient Oral Microbiome Reconstruction.Molecular ecology resources · 2026Article
- Target Capture of Ancient Shell DNA Enables Phylogenetic Reconstruction of Deep-Sea Molluscs.Molecular ecology resources · 2026Article
- Population genomics, demography, and circum-Baltic connectivity of Early Medieval southwestern Finland.Genome biology · 2026Article
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- Ancient genomes from Ladakh reveal 2800-year-old admixture between Tibetans and South Asians.Science advances · 2026Article
- Past subarctic marine food web shifts recovered by sedaDNA and network analysis.Scientific reports · 2026Article
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- Ground squirrel coprolites preserve complex archives of ancient environmental DNA over 700,000 years.Nature communications · 2026Article
- FINE-EM-seq: a rapid isothermal amplification method enabling comprehensive methylome profiling of zebrafish early embryos.Cell insight · 2026Article
- Bioarchaeological analysis illustrates the life of a 16th-century Sámi individual from Kitka, Kuusamo, northern Finland.BMC genomics · 2026Article
- Sedimentary Metagenomics Reveal Avian Community Transitions From the Last Glacial Maximum to the Holocene.Ecology and evolution · 2026Article
- The genomic history of Streptococcus mutans from the Mesolithic until modern times.Genome biology · 2026Article
- Multidisciplinary analyses and ancient DNA reveal social inequality and mobility in the Central Plains during the Eastern Zhou period in China.Nature human behaviour · 2026Article
- Ancient genomes from Ladakh reveal 2800-year-old mixture between Tibetans and South Asians.bioRxiv : the preprint server for biology · 2026Article
- DNA virus-host patterns in lake and marine environments over the last glacial cycle.The ISME journal · 2026Article
- Continuity and discontinuity in Native American populations: Insights from ancient and modern mitochondrial DNA.Genetics and molecular biology · 2026Article
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- Archaeogenetics reconstructs demography and extreme parental consanguinity in a Bronze Age community from Southern Italy.Communications biology · 2025Article
- Wet Lab Protocols Matter: Choice of DNA Extraction and Library Preparation Protocols Bias Ancient Oral Microbiome Recovery.Molecular ecology resources · 2025Article
76 more citing papers are in PubMed but not listed here.
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Authors and funding
9 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
DNA library preparation for high-throughput sequencing of genomic DNA usually involves ligation of adapters to double-stranded DNA fragments. However, for highly degraded DNA, especially ancient DNA, library preparation has been found to be more efficient if each of the two DNA strands are converted into library molecules separately. We present a new method for single-stranded library preparation, ssDNA2.0, which is based on single-stranded DNA ligation with T4 DNA ligase utilizing a splinter oligonucleotide with a stretch of random bases hybridized to a 3΄ biotinylated donor oligonucleotide. A thorough evaluation of this ligation scheme shows that single-stranded DNA can be ligated to adapter oligonucleotides in higher concentration than with CircLigase (an RNA ligase that was previously chosen for end-to-end ligation in single-stranded library preparation) and that biases in ligation can be minimized when choosing splinters with 7 or 8 random nucleotides. We show that ssDNA2.0 tolerates higher quantities of input DNA than CircLigase-based library preparation, is less costly and better compatible with automation. We also provide an in-depth comparison of library preparation methods on degraded DNA from various sources. Most strikingly, we find that single-stranded library preparation increases library yields from tissues stored in formalin for many years by several orders of magnitude.
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