A long-hidden prevalent PCR artifact and its application in DNA library screening
Jiang, X.; Jorgensen, T. S.; Wang, F.; Lien, A.; Gren, T.; Munro, L. J.; Zhang, H.; Weber, T.
Show abstract
PCR artifacts can lead to aberrant DNA products and confound result interpretation, posing significant challenges in research and diagnostics. Traditionally, it has been blamed to misannealing of primers on unintended sites. However, by sequencing, we find that most PCR byproducts and artifacts actually have both primers bound correctly but lack a middle part of the target, a phenomenon we named as PCR leaping. The leaping occurs at random positions, is independent of sequence repeats and requires a single piece template, indicating an unprecedented mechanism. It is observed with various polymerases, template qualities, GC contents, and PCR programs, thus necessitating a reset of our PCR understanding and careful consideration in future studies, for example in CRISPR editing validations. On the other hand, it provides a simple way of DNA end pairing, based on which we develop a high throughput DNA library screening method for natural product discovery at substantially reduced cost.
Matching journals
The top 5 journals account for 50% of the predicted probability mass.
Similar papers in this journal
- Comparative analysis of methodologies for detecting extrachromosomal circular DNA 95%
- Olivar: automated variant aware primer design for multiplex tiled amplicon sequencing of pathogens 95%
- Genome-wide functional screens enable the prediction of high activity CRISPR-Cas9 and -Cas12a guides in Yarrowia lipolytica 95%
Similar papers in this journal
- Rgen-Seq For Highly Sensitive Amplification-Free Screen Of Off-Target Sites Of Gene Editors 95%
- CRISPR-Based Transcriptional Activation Tool for Silent Genes in Filamentous Fungi 94%
- Identifying the essential genes of Mycobacterium avium subsp. hominissuis with Tn-Seq using a rank-based filter procedure. 94%
Similar papers in this journal
- High-resolution mapping of Sigma Factor DNA Binding Sequences using Artificial Promoters, RNA Aptamers and Deep Sequencing 95%
- ConSeqUMI, an error-free nanopore sequencing pipeline to identify and extract individual nucleic acid molecules from heterogeneous samples 95%
- Activating natural product synthesis using CRISPR interference and activation systems in Streptomyces 95%
Similar papers in this journal
- A Genome-Phenome Association study in native microbiomes identifies a mechanism for cytosine modification in DNA and RNA 95%
- RAY: CRISPR diagnostic for rapid and accurate detection of SARS-CoV2 variants on a paper strip 94%
- Linking plasmid-based beta-lactamases to their bacterial hosts using single-cell fusion PCR 94%
Similar papers in this journal
- Simple and efficient modification of Golden Gate design standards and parts using oligo stitching 96%
- Highly parallelized construction of DNA from low-cost oligonucleotide mixtures using Data-optimized Assembly Design and Golden Gate 95%
- StreptoCAD: An open-source software toolbox automating genome engineering workflows in streptomycetes 95%
"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.