Journal: bioRxiv
Article Title: Using carrier DNA in ultra-low input library preparations for next-generation sequencing
doi: 10.64898/2026.01.26.701515
Figure Lengend Snippet: Lambda DNA and ATCC mock community DNA were sheared, mixed in a 1:1 ratio, and diluted to yield 0.05 pg to 50 pg inputs. ( A ) For each input level, total read depth is shown (brown columns) together with on-target reads (mapping to Lambda and ATCC genome sequences) in gray. The number of reads mapping to Lambda DNA (purple) and ATCC DNA references (blue) are shown. Error bars indicate standard deviation associated with three replicates. The average number of cycles of PCR used for amplification is shown at the top in brown numbering. The average measured mapping ratio of Lambda DNA to ATCC DNA is shown below the PCR cycle number in purple lettering. ( B ) The relative abundance of reads mapping to each of the 10 ATCC mock community fungal genomes are shown. Results are shown for libraries with 0.05 to 50 pg DNA input amounts. The dark gray boxes indicate the expected relative abundance of the input DNA (i.e., 10% relative abundance for each organism). Error bars indicate the standard deviation associated with three technical replicates. ( C ) Distribution of ‘Ideal Scores’ for ATCC mock community sequences across experiments. Ideal scores are a univariate metric measuring the summed divergence of observed microbial communities from expected microbial communities. The metric ranges from 0 (perfect fit) to 200, with lower numbers indicating better matching between observed and expected relative abundances across all taxa. An ANOVA was performed to determine if input DNA concentrations affected the observed microbial community structure. Error bars indicate the standard deviation associated with three replicates.
Article Snippet: Three DNA templates were used for analysis in this study, including purified Enterobacteria phage lambda DNA (Lambda DNA; Oxford Nanopore Technologies, EXP-CTL001), ZymoBIOMICS Microbial Community DNA Standard (Zymo Research, D6306), and Mycobiome Genomic DNA Mix (ATCC, MSA-1010).
Techniques: Lambda DNA Preparation, Standard Deviation, Amplification
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