Skip to main content
Log in

Genotyping DNA isolated from UV irradiated human bloodstains using whole genome amplification

  • Original Article
  • Published:
Molecular Biology Reports Aims and scope Submit manuscript

Abstract

Analysis of DNA polymorphisms are the primary technique used for personal identification in forensic cases. However, DNA samples collected as evidence from crime scenes are usually degraded by environmental, physical, and chemical factors, which may interfere with the PCR analysis and, consequently, personal identification. Whole genome amplification (WGA) is a useful method to amplify genomic DNA from samples containing low quantity and poor quality of DNA, and it approach that shows promise to overcome the limited small fragments based upon random fragmentation by universal priming sites. In this study, we describe the use of WGA to genotype 15 short tandem repeat (STR) loci from dried blood samples irradiated with different types of ultraviolet (UV) light (UVA, UVB, and UVC). The result showed that UVC was the most damaging to DNA, followed by UVB and UVA. Samples exposed to UVA could be genotyped for all STR loci with or without WGA. For UVB and UVC irradiated blood samples, a greater number of STR loci could be genotyped after WGA. Although it hard to amplified a few higher molecular weight alleles, overall, the WGA method was useful in genotyping template DNA of poor quality but low quantity.

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.

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Kayser M (2015) Forensic DNA phenotyping: predicting human appearance from crime scene material for investigative purposes. Forensic Sci Int Genet 18:33–48

    Article  CAS  Google Scholar 

  2. Friedberg EC (2003) DNA damage and repair. Nature 421:436–440

    Article  Google Scholar 

  3. Hall A, Ballantyne J (2004) Characterization of UVC-induced DNA damage in bloodstains: forensic implications. Anal Bioanal Chem 380:72–83

    Article  CAS  Google Scholar 

  4. Poepping C, Beck SE, Wright H, Linden KG (2014) Evaluation of DNA damage reversal during medium-pressure UV disinfection. Water Res 56:181–189

    Article  CAS  Google Scholar 

  5. Vaisman A, Woodgate R (2017) Translesion DNA polymerases in eukaryotes: what makes them tick? Crit Rev Biochem Mol Biol 52(3):274–303

    Article  CAS  Google Scholar 

  6. Diegoli TM, Farr M, Cromartie C, Coble MD, Bille TW (2012) An optimized protocol for forensic application of the PreCR™ Repair Mix to multiplex STR amplification of UV-damaged DNA. Forensic Sci Int Genet 6(4):498–503

    Article  CAS  Google Scholar 

  7. Hall A, Sims LM, Ballantyne J (2014) Assessment of DNA damage induced by terrestrial UV irradiation of dried bloodstains: forensic implications. Forensic Sci Int Genet 8(1):24–32

    Article  CAS  Google Scholar 

  8. Rinke J, Schäfer V, Schmidt M, Ziermann J, Kohlmann A, Hochhaus A, Ernst T (2013) Genotyping of 25 leukemia-associated genes in a single work flow by next-generation sequencing technology with low amounts of input template DNA. Clin Chem 59(8):1238–1250

    Article  CAS  Google Scholar 

  9. Han T, Chang CW, Kwekel JC, Chen Y, Ge Y, Martinez-Murillo F, Roscoe D, Težak Z, Philip R, Bijwaard K, Fuscoe JC (2012) Characterization of whole genome amplified (WGA) DNA for use in genotyping assay development. BMC Genom 13(217):1–15

    Google Scholar 

  10. Huang L, Ma F, Chapman A, Lu S, Xie XS (2015) Single-cell whole-genome amplification and sequencing: methodology and applications. Annu Rev Genomics Hum Genet 16:79–102

    Article  CAS  Google Scholar 

  11. Borgström E, Paterlini M, Mold JE, Frisen J, Lundeberg J (2017) Comparison of whole genome amplification techniques for human single cell exome sequencing. PLoS ONE 12(2):e0171566

    Article  Google Scholar 

  12. Lindahl T (1993) Instability and decay of the primary structure of DNA. Nature 362:709–715

    Article  CAS  Google Scholar 

  13. Robertson JM, Dineen SM, Scott KA, Lucyshyn J, Saeed M, Murphy DL, Schweighardt AJ, Meiklejohn KA (2014) Assessing PreCR™ repair enzymes for restoration of STR profiles from artificially degraded DNA for human identification. Forensic Sci Int Genet 12:168–180

    Article  CAS  Google Scholar 

  14. Mitchell D, Willerslev E, Hansen A (2005) Damage and repair of ancient DNA. Mutat Res 571:265–276

    Article  CAS  Google Scholar 

  15. Tate CM, Nuñez AN, Goldstein CA, Gomes I, Robertson JM, Kavlick MF, Budowle B (2012) Evaluation of circular DNA substrates for whole genome amplification prior to forensic analysis. Forensic Sci Int Genet 6(2):185–190

    Article  CAS  Google Scholar 

  16. Kroneis T, El-Heliebi A (2015) Quality control of isothermal amplified DNA based on short tandem repeat analysis. Methods Mol Biol 1347:129–140

    Article  CAS  Google Scholar 

  17. Lee JC, Tsai LC, Lai PY, Lee CC, Lin CY, Huang TY, Linacre A, Hsieh HM (2012) Evaluating the performance of whole genome amplification for use in low template DNA typing. Med Sci Law 52(4):223–228

    Article  Google Scholar 

  18. Nishikawa Y, Hosokawa M, Maruyama T, Yamagishi K, Mori T, Takeyama H (2015) Monodisperse picoliter droplets for low-bias and contamination-free reactions in single-cell whole genome amplification. PLoS ONE 10(9):e0138733

    Article  Google Scholar 

Download references

Acknowledgements

We thank Mr. Eiji Isobe for assistance with the samples irradiation.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Jian Tie.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

Approval for this study was granted by the Ethics Committee of Nihon University School of Medicine and National Research Committee. All procedures performed in studies involving human participants were in accordance with the ethical standards of this Ethics Committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all participants included in the study.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Uchigasaki, S., Tie, J., Sobashima, E. et al. Genotyping DNA isolated from UV irradiated human bloodstains using whole genome amplification. Mol Biol Rep 45, 925–929 (2018). https://doi.org/10.1007/s11033-018-4240-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11033-018-4240-6

Keywords

Navigation