Skip to main content
Log in

PCR-based evaluation of sequence specificity of DNA fragmentation by ultrasound

  • Genomics. Transcriptomics
  • Published:
Molecular Biology Aims and scope Submit manuscript

Abstract

Ultrasonic fragmentation, which is a simple and convenient method for the mechanical degradation of DNA, is widely used in modern genome studies as one of the sample preparation steps. It has been recently found that the DNA breaks occur more often in the regions containing 5'-CG-3' dinucleotides. We studied the influence of the 5'-CG-3' dinucleotides on the efficiency of the 28S rRNA gene amplification during PCR with sonicated DNA of Mantis religiosa. It was shown that the amplification rate depends on the template length and the number of 5'-CG-3' dinucleotides. Amplification of the DNA regions with a higher 5'-CG-3' density is less efficient because of their higher sensitivity to ultrasound. The amount of the amplified DNA templates is inversely proportional to the 5'-CG-3'number.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Ansorge W.J. 2009. Next-generation DNA sequencing techniques. N. Biotechnol. 25, 195–203.

    Article  CAS  PubMed  Google Scholar 

  2. Mann T.L., Krull U.J. 2004. The application of ultrasound as a rapid method to provide DNA fragments suitable for detection by DNA biosensors. Biosens. Bioelectron. 20, 945–955.

    Article  CAS  PubMed  Google Scholar 

  3. Elsner H.I., Lindblad E.B. 1989. Ultrasonic degradation of DNA. DNA. 8, 697–701.

    Article  CAS  PubMed  Google Scholar 

  4. Taub M.A., Bravo H.C., Irizarry R.A. 2010. Overcoming bias and systematic errors in next generation sequencing data. Genome Med. 2, 87.

    Article  PubMed  PubMed Central  Google Scholar 

  5. Schwartz S., Oren R., Ast G. 2011. Detection and removal of biases in the analysis of next-generation sequencing reads. PLoS ONE. 6, e16685.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Benjamini Y., Speed T.P. 2012. Summarizing and correcting the GC content bias in high-throughput sequencing. Nucleic Acids Res. 40, e72.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Guo Y., Li J., Li Ch., Long J., Samuels D.C., Shyr Y. 2012. The effect of strand bias in Illumina short-read sequencing data. BMC Genomics. 13, 666.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Chen Y.-C., Liu T., Yu C.-H., Chiang T.-Y., Hwang C.-C. 2013. Effects of GC bias in next-generation-sequencing data on de novo genome assembly. PLoS ONE. 8, e62856.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Poptsova M.S., Il’icheva I.A., Nechipurenko D.Yu., Panchenko L.A., Khodikov M.V., Oparina N.Y., Polozov R.V., Nechipurenko Yu.D., Grokhovsky S.L. 2014. Non-random DNA fragmentation in next-generation sequencing. Sci. Reports. 4, 4532.

    Google Scholar 

  10. Packer M.J., Dauncey M.P., Hunter C.A. 2000. Sequence-dependent DNA structure: Tetranucleotide conformational maps. J. Mol. Biol. 295, 85–103.

    Article  CAS  PubMed  Google Scholar 

  11. Grokhovsky S.L. 2006. Specificity of DNA cleavage by ultrasound. Mol. Biol. (Moscow). 40 (2), 275–283.

    Article  Google Scholar 

  12. Nechipurenko Yu.D., Golovkin M.V., Nechipurenko D.Yu., Il’icheva I.A., Panchenko L.A., Polozov R.V., Grokhovsky S.L. 2009. Characteristic features of DNA cleavage by ultrasound. Zh. Strukt. Khim. 50, 1045–1052.

    Google Scholar 

  13. Grokhovsky S.L., Il’icheva I.A., Nechipurenko D.Yu., Golovkin M.V., Panchenko L.A., Polozov R.V., Nechipurenko Yu.D. 2011. Sequence-specific ultrasonic cleavage of DNA. Biophys. J. 100, 117–125.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Grokhovsky S.L., Il’icheva I.A., Panchenko L.A., Golovkin M.V., Nechipurenko D.Yu., Polozov R.V., Nechipurenko Yu.D. 2013. Ultrasonic cleavage of DNA in complexes with Ag(I), Cu(II), Hg(II). Biophysics (Moscow). 58 (1), 19–26.

    Article  Google Scholar 

  15. Grokhovsky S.L., Il’icheva I.A., Nechipurenko D.Yu., Panchenko L.A., Polozov R.V., Nechipurenko Yu.D. 2008. Heterogeneity of dounble-stranded DNA local structure and dynamics: Ultrasound studies. Biofizika. 53, 417–425.

    Google Scholar 

  16. Nechipurenko D.Yu., Il’icheva I.A., Khodikov M.V., Poptsova M.S., Nechipurenko Yu.D., Grokhovsky S.L. 2014. Modeling of mechanochemical DNA cleavage by the action of ultrasound. Biophysics (Moscow). 59 (6), 861–868.

    Article  CAS  Google Scholar 

  17. Nechipurenko Yu.D., Nechipurenko D.Yu, Il’icheva I.A., Golovkin M.V., Panchenko L.A., Polozov R.V., Grokhovsky S.L. 2010. Conformational dynamic properties of DNA and approaches to physical genome mapping. Komp’yut. Issled. Model. 2, 419–428.

    Google Scholar 

  18. Larguinho M., Santos H.M., Doria G., Scholz H., Baptista P.V., Capelo J.L. 2010. Development of a fast and efficient ultrasonic-based strategy for DNA fragmentation. Talanta. 81, 881–886.

    Article  CAS  PubMed  Google Scholar 

  19. Hagerman P.J. 1981. Investigation of the flexibility of DNA using transient electric birefringence. Biopolymers. 20, 1503–1535.

    Article  CAS  PubMed  Google Scholar 

  20. Golenberg E.M., Bickel A., Weihs P. 1996. Effect of highly fragmented DNA on PCR. Nucleic Acids Res. 24, 5026–5033.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Garafutdinov R.R., Nagaev N.R., Sakhabutdinova A.R., Chemeris A.V. 2015. Authenticity, integrity, and accessibility of ancient DNA. Vestn. Bashkir. Gos. Univ. 20, 432–439.

    Google Scholar 

  22. http://www.ncbi.nlm.nih.gov/nuccore/

  23. http://eu.idtdna.com/analyzer/Applications/OligoAnalyzer/

  24. Koshikawa S., Miyazaki S., Cornette R., Matsumoto T., Miura T. 2008. Genome size of termites (Insecta, Dictyoptera, Isoptera) and wood roaches (Insecta, Dictyoptera, Cryptocercidae). Naturwissenschaften. 95, 859–867.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to R. R. Garafutdinov.

Additional information

Original Russian Text © R.R. Garafutdinov, A.A. Galimova, A.R. Sakhabutdinova, A.V. Chemeris, 2016, published in Molekulyarnaya Biologiya, 2016, Vol. 50, No. 2, pp. 272–278.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Garafutdinov, R.R., Galimova, A.A., Sakhabutdinova, A.R. et al. PCR-based evaluation of sequence specificity of DNA fragmentation by ultrasound. Mol Biol 50, 236–241 (2016). https://doi.org/10.1134/S0026893316020059

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1134/S0026893316020059

Keywords

Navigation