The use of iconPCR for 16S library preparation improves data quality and workflow
Patel, P.
Show abstract
Polymerase Chain Reaction (PCR) is a cornerstone of contemporary biological research, enabling the amplification of specific DNA sequences for various applications. However, suboptimal cycling conditions often undermine its efficacy, which can generate chimeric products, exacerbate PCR duplication rates, and skew species representation in metabarcoding experiments due to the preferential amplification of dominant populations. To address these limitations, we present iconPCR--Individually Controlled PCR--a novel technology that allows each reaction in a 96-well plate to be cycled independently and programmatically. By setting a predefined fluorescence threshold, iconPCR ensures that all Next Generation Sequencing (NGS) libraries are amplified to equivalent levels, thereby eliminating the risks of over- or under-amplification through a process known as "Auto-Normalization." In this study, we applied iconPCR to generate V3, V4, and full-length 16S rRNA gene libraries using Avidite sequencing. The V1-V9 variable region libraries were also evaluated with HiFi sequencing, which significantly demonstrated its potential to improve microbial community analysis accuracy and reliability.
Matching journals
The top 5 journals account for 50% of the predicted probability mass.
Similar papers in this journal
- Benchmarking long-read sequencing strategies for obtaining ASV-resolved rRNA operons from environmental microeukaryotes 97%
- High molecular weight DNA extraction strategies for long-read sequencing of complex metagenomes 97%
- Evaluation of primer pairs for microbiome profiling across a food chain from soils to humans within the One Health framework 96%
Similar papers in this journal
- Evaluation of the effects of library preparation procedure and sample characteristics on the accuracy of metagenomic profiles 97%
- Measurement error and resolution in quantitative stable isotope probing: implications for experimental design 95%
- Evaluating and Improving SSU rRNA PCR Primer Coverage for Bacteria, Archaea, and Eukaryotes Using Metagenomes from Global Ocean Surveys 94%
Similar papers in this journal
- An improved hgcAB primer set and direct high throughput sequencing expand Hg-methylator diversity in nature 96%
- Interpretations of microbial community studies are biased by the selected 16S rRNA gene amplicon sequencing pipeline. 96%
- Ensuring that fundamentals of quantitative microbiology are reflected in microbial diversity analyses based on next-generation sequencing 95%
Similar papers in this journal
- Detecting Flavobacterial Fish Pathogens in the Environment Using High-Throughput Community Analysis 95%
- intI1 primer selection for class 1 integron integrase gene and transcript quantification - validation and application for monitoring intl1 gene abundance within septic tanks in Thailand 94%
- Seafloor incubation experiment with deep-sea hydrothermal vent fluid reveals effect of pressureand lag time on autotrophic microbial communities 94%
Similar papers in this journal
- Library Preparation and Sequencing Platform Introduce Bias in Metagenomic-Based Characterizations of Microbiomes 95%
- rRNA Operon Improves Species-Level Classification of Bacteria and Microbial Community Analysis Compared to 16S rRNA 94%
- Machine-learning based detection of adventitious microbes in T-cell therapy cultures using long read sequencing 94%
"Similar papers" are the closest papers from that journal in the model's embedding space. They show what the match is built on, but the ranking comes mostly from a classifier over the whole training set, not from these examples alone.