Document Type
Article
Journal Title
Nucleic Acids Research
Publication Date
4-12-2024
Volume
52
Abstract
Custom oligonucleotides (oligos) are widely used reagents in biomedical research. Some common applications of oligos include polymerase chain reaction (PCR), sequencing, hybridization, microarray, and library construction. The reliability of oligos in such applications depends on their purity and specificity. Here, we report that commercially available oligos are frequently contaminated with nonspecific sequences (i.e. other unrelated oligonucleotides). Most of the oligos that we designed to amplify clustered regularly interspersed palindromic repeats (CRISPR) guide sequences contained nonspecific CRISPR guides. These contaminants were detected in research-grade oligos procured from eight commercial oligo-suppliers located in three different geographic regions of the world. Deep sequencing of some of the oligos revealed a variety of contaminants. Given the wide range of applications of oligos, the impact of oligo cross-contamination varies greatly depending on the field and the experimental method. Incorporating appropriate control experiments in research design can help ensure that the quality of oligo reagents meets the intended purpose. This can also minimize risk depending on the purposes for which the oligos are used.
MeSH Headings
Base Sequence, Clustered Regularly Interspaced Short Palindromic Repeats, Oligonucleotides, Genetic Techniques, Drug Contamination, Indicators and Reagents, Industry
DOI Link
ISSN
1362-4962
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License
Recommended Citation
Arakawa, Hiroshi; Miura, Hiromi; Quadros, Rolen M.; Ohtsuka, Masato; and Gurumurthy, Channabasavaiah B., "Cross-Contamination of CRISPR Guides and Other Unrelated Nucleotide Sequences Among Commercial Oligonucleotides" (2024). Journal Articles: Genetics, Cell Biology & Anatomy. 56.
https://digitalcommons.unmc.edu/com_gcba_articles/56