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Fast Masking of Repeated Primer Binding Sites in Eukaryotic Genomes

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PCR Primer Design

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1275))

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Abstract

In this article we describe the working principle and a list of practical applications for GenomeMasker—a program that finds and masks all repeated DNA motifs in fully sequenced genomes. The GenomeMasker exhaustively finds and masks all repeated DNA motifs in studied genomes. The software is optimized for PCR primer design. The algorithm is designed for high-throughput work, allowing masking of large DNA regions, even entire eukaryotic genomes. Additionally, the software is able to predict all alternative PCR products from studied genomes for thousands of candidate PCR primer pairs.

Practical applications of the GenomeMasker are shown for command-line version of the GenomeMasker, which can be downloaded from http://bioinfo.ut.ee/download/. Graphical Web interfaces with limited options are available at http://bioinfo.ut.ee/genometester/ and http://bioinfo.ut.ee/snpmasker/.

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References

  1. Dawson E, Abecasis GR, Bumpstead S et al (2002) A first-generation linkage disequilibrium map of human chromosome 22. Nature 418:544–548

    Article  CAS  PubMed  Google Scholar 

  2. Rouchka EC, Khalyfa A, Cooper NG (2005) MPrime: efficient large scale multiple primer and oligonucleotide design for customized gene microarrays. BMC Bioinformatics 6:175

    Article  PubMed Central  PubMed  Google Scholar 

  3. Xu D, Li G, Wu L et al (2002) PRIMEGENS: robust and efficient design of gene-specific probes for microarray analysis. Bioinformatics 18:1432–1437

    Article  CAS  PubMed  Google Scholar 

  4. Weckx S, De Rijk P, Van Broeckhoven C et al (2005) SNPbox: a modular software package for large-scale primer design. Bioinformatics 21:385–387

    Article  CAS  PubMed  Google Scholar 

  5. Zhang Z, Schwartz S, Wagner L et al (2000) A greedy algorithm for aligning DNA sequences. J Comput Biol 7:203–214

    Article  CAS  PubMed  Google Scholar 

  6. Ning Z, Cox AJ, Mullikin JC (2001) SSAHA: a fast search method for large DNA databases. Genome Res 11:1725–1729

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  7. Kent WJ (2002) BLAT–the BLAST-like alignment tool. Genome Res 12:656–664

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  8. Benson G (1999) Tandem repeats finder: a program to analyze DNA sequences. Nucleic Acids Res 27:573–580

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  9. Altschul SF, Madden TL, Schaffer AA et al (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  10. Rouillard JM, Zuker M, Gulari E (2003) OligoArray 2.0: design of oligonucleotide probes for DNA microarrays using a thermodynamic approach. Nucleic Acids Res 31:3057–3062

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  11. Van Hijum SA, De Jong A, Buist G et al (2003) UniFrag and GenomePrimer: selection of primers for genome-wide production of unique amplicons. Bioinformatics 19:1580–1582

    Article  PubMed  Google Scholar 

  12. Untergrasser A, Cutcutache I, Koressaar T et al (2012) Primer3 – new capabilities and interfaces. Nucleic Acids Res 40:e115

    Article  Google Scholar 

  13. Breslauer KJ, Frank R, Blocker H et al (1986) Predicting DNA duplex stability from the base sequence. Proc Natl Acad Sci U S A 83:3746–3750

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  14. SantaLucia J Jr (1998) A unified view of polymer, dumbbell, and oligonucleotide DNA nearest-neighbor thermodynamics. Proc Natl Acad Sci U S A 95:1460–1465

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  15. Owczarzy R, You Y, Moreira BG et al (2004) Effects of sodium ions on DNA duplex oligomers: improved predictions of melting temperatures. Biochemistry 43:3537–3554

    Article  CAS  PubMed  Google Scholar 

  16. Von Ahsen N, Wittwer CT, Schutz E (2001) Oligonucleotide melting temperatures under PCR conditions: nearest-neighbor corrections for Mg(2+), deoxynucleotide triphosphate, and dimethyl sulfoxide concentrations with comparison to alternative empirical formulas. Clin Chem 47:1956–1961

    Google Scholar 

  17. Kaplinski L, Andreson R, Puurand T et al (2005) MultiPLX: automatic grouping and evaluation of PCR primers. Bioinformatics 21:1701–1702

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

The development of GenomeMasker package was supported by the Estonian Ministry of Education and Research grant 0182649s04 and grant EU19730 from Enterprise Estonia. Development of primer design software in our group has been funded by Centre of Excellence in Genomics at Estonian Biocentre (EU European Regional Development Fund). The authors thank Katre Palm for a valuable help with English grammar.

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Correspondence to Maido Remm .

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Andreson, R., Kaplinski, L., Remm, M. (2015). Fast Masking of Repeated Primer Binding Sites in Eukaryotic Genomes. In: Basu, C. (eds) PCR Primer Design. Methods in Molecular Biology, vol 1275. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2365-6_1

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  • DOI: https://doi.org/10.1007/978-1-4939-2365-6_1

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2364-9

  • Online ISBN: 978-1-4939-2365-6

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