Abstract
Previous studies on DNA damage and repair have involved in vitro laboratory procedures that induce a single type of lesion in naked templates. Although repair of singular, sequestered types of DNA damage has shown some success, forensic and ancient specimens likely contain a number of different types of lesions. This study sought to (1) develop protocols to damage DNA in its native state, (2) generate a pool of candidate samples for repair that more likely emulate authentic forensic samples, and (3) assess the ability of the PreCRTM Repair Mix to repair the resultant lesions. Complexed, native DNA is more difficult to damage than naked DNA. Modified procedures included the use of higher concentrations and longer exposure times. Three types of samples, those that demonstrated damage based on short tandem repeat (STR) profile signals, were selected for repair experiments: environmentally damaged bloodstains, bleach-damaged whole blood, and human skeletal remains. Results showed trends of improved performance of STR profiling of bleach-damaged DNA. However, the repair assay did not improve DNA profiles from environmentally damaged bloodstains or bone, and in some cases resulted in lower RFU values for STR alleles. The extensive spectrum of DNA damage and myriad combinations of lesions that can be present in forensic samples appears to pose a challenge for the in vitro PreCRTM assay. The data suggest that the use of PreCR in casework should be considered with caution due to the assay’s varied results.
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Acknowledgement
This project was supported by the National Institute of Justice, Office of Justice Programs, U.S. Department of Justice (Award No. 2010-DN-BX-K227). The opinions, findings, and conclusions or recommendations expressed in this publication are those of the authors and do not necessarily reflect those of the U.S. Department of Justice.
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Supplementary Table 1
Principal sources of DNA damage and major types of DNA lesions. (XLSX 11 kb)
Supplementary Figure 1
Average peak height from damaged DNA in whole human blood after immersion in a 50 % Clorox® bleach solution (3 % NaOCl) and average peak height after treatment w/PreCR™ Repair Mix (manufacturer-recommended protocol). n = 40 blood samples (each repaired in duplicate, for a total of 80 repair reactions) (GIF 235 kb)
Supplementary Figure 2
Comparison of average peak heights from bleach-damaged DNA with and without treatment with PreCR™ Repair Mix (manufacturer vs. modified protocol). n = 25 blood samples (each repaired in duplicate, for a total of 50 repair reactions) (GIF 313 kb)
Supplementary Figure 3
Average peak heights of environmentally damaged DNA in bloodstains with and without treatment with PreCR™ Repair Mix (manufacturer-recommended protocol). n = 75 bloodstains (each repaired in duplicate, for a total of 150 repair reactions) (GIF 211 kb)
Supplementary Figure 4
Average peak heights of environmentally damaged DNA in bloodstains with and without PreCR™ Repair Mix treatment (manufacturer-recommended protocol vs. modified protocol). n = 30 bloodstains (each repaired in duplicate, for a total of 60 repair reactions) (GIF 349 kb)
Supplementary Figure 5
Average peak heights of damaged DNA from contemporary human skeletal remains with and without PreCR™ Repair Mix treatment (manufacturer-recommended protocol). n = 50 bone samples (each repaired in duplicate, for a total of 100 repair reactions) (GIF 255 kb)
Supplementary Figure 6
Average peak heights of damaged DNA from contemporary human skeletal remains with and without PreCR™ Repair Mix treatment (manufacturer-recommended protocol vs. modified protocol). n = 30 bone samples (each repaired in duplicate, for a total of 60 repair reactions) (GIF 270 kb)
Supplementary Figure 7
Average peak heights of damaged DNA from historical human skeletal remains with and without PreCR™ Repair Mix treatment (modified protocol). Bones were 120 years old. (n = 20) (GIF 236 kb)
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Ambers, A., Turnbough, M., Benjamin, R. et al. Assessment of the role of DNA repair in damaged forensic samples. Int J Legal Med 128, 913–921 (2014). https://doi.org/10.1007/s00414-014-1003-3
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DOI: https://doi.org/10.1007/s00414-014-1003-3