TY - JOUR
T1 - Assessment of the role of DNA repair in damaged forensic samples
AU - Ambers, Angie
AU - Turnbough, Meredith
AU - Benjamin, Robert
AU - King, Jonathan
AU - Budowle, Bruce
N1 - Funding Information:
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.
Publisher Copyright:
© 2014, Springer-Verlag Berlin Heidelberg.
PY - 2014/11
Y1 - 2014/11
N2 - 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.
AB - 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.
KW - DNA damage
KW - DNA repair
KW - Human skeletal remains
KW - PreCR™ Repair Mix
UR - http://www.scopus.com/inward/record.url?scp=84934267774&partnerID=8YFLogxK
U2 - 10.1007/s00414-014-1003-3
DO - 10.1007/s00414-014-1003-3
M3 - Article
C2 - 24792635
AN - SCOPUS:84934267774
SN - 0937-9827
VL - 128
SP - 913
EP - 921
JO - International Journal of Legal Medicine
JF - International Journal of Legal Medicine
IS - 6
ER -