Year: 2023
Pages: 87-95
Number: Volume 15, issue 2
Type: scientific article
DOI: https://doi.org/10.31301/2221-6197.bmcs.2023-12
Topic: Articls
Authors: Garafutdinov R.R.!, Galimova A.A., Sakhabutdinova A.R.!, Chemeris A.V.
Fragmentation of DNA by ultrasound is the most optimal method of artificial degradation of DNA molecules. It requires no reagents and makes it possible to obtain "pure" DNA samples for further use. However, despite an active usage of ultrasonic fragmentation in studies, a number of issues related to the applicability of mechanically degraded DNA for standard molecular biological operations remain poorly studied. This report presents data on the effect of the size of DNA molecules (on DNA amplicons) and the composition of aqueous solutions on the efficiency of DNA destruction by ultrasound. It has been shown that the rate of DNA fragmentation positively correlates with an increase in the length of molecules. In solutions with high ionic strength, the rate of fragmentation increases dramatically. The presence of organic solvents that do not affect hybridization slightly reduces the rate of ultrasonic destruction. With an increase in the length of DNA molecules, the influence of denaturing agents changes and leads to an increase in the rate of fragmentation. dsDNA molecules smaller than one persistent length (about 150 bp) are not fragmented.
DNA, amplicons, ultrasound, mechanical fragmentation, persistent length
1. Ansorge W.J. Next-generation DNA sequencing techniques. Nature Biotechnol. 2009. V. 25. P. 195-203. doi: 10.1016/j.nbt.2008.12.009
2. Bikbulatova S.M., Chemeris D.A., Nikonorov Yu.M., Mashkov O.I., Garafutdinov R.R., Chemeris A.V., Vakhitov V.A. Methods for detection of results of realtime polymerase chain reaction. Vestn. Bashk. University. 2012. V. 17(1). P. 59-67. (In Russian)
3. Chemeris D.A., Magdanov E.G., Mashkov O.I., Garafutdinov R.R., Chemeris A.V. Delayed or hot-start PCR. Biomics. 2011. V. 2(1). P. 1-8. (In Russian)
4. Chen Y.-C., Liu T., Yu C.-H., Chiang T.-Y., Hwang C.-C. Effects of GC bias in next-generation-sequencing data on de novo genome assembly. PLoS ONE. 2013. V. 8. e62856. doi: 10.1371/journal.pone.0062856
5. Elsner H.I., Lindblad E.B. Ultrasonic degradation of DNA. DNA. 1989. V. 8. P. 697-701. doi: 10.1089/dna.1989.8.697
6. Garafutdinov R.R., Galimova А.А., Sakhabutdinova А.R. Polymerase chain reaction with nearby primers. Anal. Biochem. 2017. V. 518. P. 126-133. doi: 10.1016/j.ab.2016.11.017
7. Garafutdinov R.R., Galimova A.A., Sakhabutdinova A.R., Chemeris A.V. PCR-based evaluation of sequence specificity of DNA fragmentation by ultrasound. Mol. Biol. (Mosk). 2016. V. 50(2). P. 272-278. doi: 10.7868/S0026898416020051
8. Garafutdinov R.R., Galimova А.А., Sakhabutdinova А.R. The influence of CpG (5'-d(CpG)-3' dinucleotides) methylation on ultrasonic DNA fragmentation. J. Biomol. Struct. Dyn. 2019. V. 37. P. 3877-3886. doi: 10.1080/07391102.2018.1533888
9. Garafutdinov R.R., Galimova A.A., Sakhabutdinova A.R., Vakhitov V.A., Chemeris A.V. DNA amplification using PCR with abutting primers. Mol. Biol. (Mosk). 2015. V. 49(4). P. 628-637. doi: 10.7868/S0026898415040059
10. Garafutdinov R.R., Nagaev N.R., Sakhabutdinova A.R., Chemeris A.V. Authenticity, preservation and availability of ancient DNA. Vestn. Bashk. University. 2015а. V. 20. P. 432-439. (In Russian)
11. Golenberg E.M., Bickel A., Weihs P. Effect of highly fragmented DNA on PCR. Nucleic Acids Res. 1996. V. 24. P. 5026-5033. doi: 10.1093/nar/24.24.5026
12. Grokhovsky S.L. Specificity of DNA cleavage by ultrasound. Mol. Biol. 2006. V. 40(2). P. 276-283. doi: 10.1134/S0026893306020142
13. Grokhovsky S.L., Il’icheva I.A., Nechipurenko D.Yu., Golovkin M.V., Panchenko L.A., Polozov R.V., Nechipurenko Yu.D. Sequence-specific ultrasonic cleavage of DNA // Biophys. J. 2011. V. 100. P. 117-125. doi: 10.1016/j.bpj.2010.10.052
14. Grokhovsky S.L., Il'icheva I.A., Nechipurenko Yu.D., Nechipurenko D.Yu., Panchenko L.A., Polozov R.V. Ultrasonic cleavage of DNA: quantitative analysis of sequence specificity. Biophysics. 2008. V. 53(3). P. 250-251.
15. Grokhovsky S.L., Il'icheva I.A., Golovkin M.V., Nechipurenko Y.D., Nechipurenko D.Y., Panchenko L.A., Polozov R.V. Ultrasonic cleavage of DNA in complexes with Ag(I), Cu(II), Hg(II). Biophysics. 2013. V. 58(1). P. 27-36. doi: 10.1134/S0006350913010065
16. Hagerman P.J. Investigation of the flexibility of DNA using transient electric birefringence. Biopolymers. 1981. V. 20. P. 1503-1535. doi: 10.1002/bip.1981.360200710
17. Larguinho M., Santos H.M., Doria G., Scholz H., Baptista P.V., Capelo J.L. Development of a fast and efficient ultrasonic-based strategy for DNA fragmentation. Talanta. 2010. V. 81. P. 881-886. doi: 10.1016/j.talanta.2010.01.032
18. Mann T.L., Krull U.J. The application of ultrasound as a rapid method to provide DNA fragments suitable for detection by DNA biosensors. Biosens. Bioelectron. 2004. V. 20. P. 945-955. doi: 10.1016/j.bios.2004.06.021
19. Mashkov O.I., Poskryakov A.V., Nikolenko A.G., Garafutdinov R.R. Determination of the botanical origin of honey using PCR. Beekeeping. 2016. No.2. P. 56-58. (In Russian)
20. Mashkov O.I., Poskryakov A.V., Nikolenko A.G., Garafutdinov R.R. Establishment of the botanical origin of honey using polymerase chain reaction. Biomics. 2016а. V. 8(2). P. 154-160. (In Russian)
21. Nechipurenko Y.D., Golovkin M.V., Il'icheva I.A., Grokhovskii S.L., Nechipurenko D.Y., Panchenko L.A., Polozov R.V. Characteristics of ultrasonic cleavage of DNA. J. Struct. Chem. 2009. V. 50 (5). P. 1007-1013. doi: 10.1007/s10947-009-0149-1
22. Packer M.J., Dauncey M.P., Hunter C.A. Sequencedependent DNA structure: tetranucleotide conformational maps. J. Mol. Biol. 2000. V. 295. P. 85-103. doi: 10.1006/jmbi.1999.3237
23. 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. Nonrandom DNA fragmentation in next-generation sequencing. Sci. Reports. 2014. V. 4. P. 4532. doi: 10.1038/srep04532
24. Sakhabutdinova A.R., Maksimova M.A., Garafutdinov R.R. Synthesis of Circular DNA Templates with T4 RNA Ligase for Rolling Circle Amplification. Mol. Biol. (Mosk). 2017. V. 51(4). P. 724-733. doi: 10.7868/S0026898417040164
25. Taub M.A., Bravo H.C., Irizarry R.A. Overcoming bias and systematic errors in next generation sequencing data. Genome Med. 2010. V. 2. P. 87. doi: 10.1186/gm208