Cited by 170Gene expression and cancer classificationAdvanced biosensing and bioanalysis techniquesGenomics and Phylogenetic Studies

Design of 240,000 orthogonal 25mer DNA barcode probes

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Xu, Qikai, Schlabach, Michael R., Hannon, Gregory J., Elledge, Stephen J.·2009·Proceedings of the National Academy of Sciences

Abstract

DNA barcodes linked to genetic features greatly facilitate screening these features in pooled formats using microarray hybridization, and new tools are needed to design large sets of barcodes to allow construction of large barcoded mammalian libraries such as shRNA libraries. Here we report a framework for designing large sets of orthogonal barcode probes. We demonstrate the utility of this framework by designing 240,000 barcode probes and testing their performance by hybridization. From the test hybridizations, we also discovered new probe design rules that significantly reduce cross-hybridization after their introduction into the framework of the algorithm. These rules should improve the performance of DNA microarray probe designs for many applications.

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  • The barcode library allows for individual selection of up to ~10^15 possible distinct files using only three unique barcodes per file.

    “which allows for individual selection of up to ~1015 possible distinct files using only three unique barcodes per file20”
  • Avoiding spurious amplification of off-target files due to crosstalk of PCR primers with incorrect barcodes or main file sequences requires careful primer design.

    “Finally, avoiding spurious amplification of off-target files due to crosstalk of PCR primers with incorrect barcodes or main file sequences requires careful primer design20.”

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Authors
Xu, Q., Schlabach, M. R., Hannon, G. J., & Elledge, S. J.
Affiliations
  • Brigham and Women's Hospital (US)
  • Howard Hughes Medical Institute (US)
Venue
Proceedings of the National Academy of Sciences
Published
2009-02-17
Volume
106
Issue
7
Pages
2289-2294
ISSN
  • 0027-8424 (ISSN-L)
  • 1091-6490
OpenAlex
W2091266578
Cited by
170
References
22
Citations by year
2026: 13 · 2025: 16 · 2024: 13 · 2023: 13 · 2022: 13 · 2021: 22 · 2020: 12 · 2019: 11 · 2018: 6 · 2017: 10 · 2016: 8 · 2015: 5 · 2014: 3 · 2013: 5 · 2012: 6
FWCI
2.68
Type
journal article
Language
en
Open access
Locations
  • Proceedings of the National Academy of Sciences (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • Cold Spring Harbor Laboratory Institutional Repository (Cold Spring Harbor Laboratory) (acceptedVersion)
Funders
Howard Hughes Medical Institute
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Gene expression and cancer classification
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Advanced biosensing and bioanalysis techniques
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Phylogenetic Studies
Keywords
DNA barcodes, probe design, microarray hybridization, cross-hybridization, shRNA library
MeSH terms
Algorithms, Electronic Data Processing, DNA, Nucleic Acid Conformation, Nucleic Acid Hybridization, RNA, Software, Temperature, DNA Probes, Oligonucleotide Probes, Sequence Analysis, DNA, Oligonucleotide Array Sequence Analysis, Gene Expression Profiling

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