An improved method for STR analysis of bloodstained denim

Larkin, A.; Harbison, SallyAnn

doi:10.1007/s004140050020

Cited by 39 publications

(17 citation statements)

References 6 publications

(10 reference statements)

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“…For IAC 410 , the optimal annealing temperature was 581C. Intercalates with DNA [48][49][50] Vaginal microorganisms Semen samples from sexual assaults DNA sequestration by single-stranded length of microbial DNA or by reduction of primer concentration by non-specific binding to microbial DNA [5] Calcium phosphate Bones Calcium ions compete with magnesium ions, which are required for the activity of the polymerase [3,33] Electrophoresis 2011, 32, 1371-1378…”

Section: Generation Of the Iacsmentioning

confidence: 99%

Development of internal amplification controls for DNA profiling with the AmpFℓSTR^® SGM Plus^® kit

et al. 2011

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DNA extracted from forensic samples can be degraded and also contain co-extracted contaminants that inhibit PCR. The effects of DNA degradation and PCR inhibition are often indistinguishable when examining a DNA profile. Two internal amplification controls (IACs) were developed to improve quality control of PCR using the AmpFℓSTR® SGM Plus® kit. The co-amplification of these controls with DNA samples was used to monitor amplification efficiency and detect PCR inhibitors. IAC fragments of 90 and 410 bp (IAC₉₀ and IAC₄₁₀) were generated from the plasmid pBR322 using tailed primers and then amplified with ROX-labelled primers. Co-amplification of IAC₉₀ and IAC₄₁₀ was performed with varying amounts of template DNA, degraded DNA and DNA contaminated with humic acid, heme and indigo dye. Both IAC₉₀ and IAC₄₁₀ were successfully amplified with human DNA without significantly affecting the quality of the DNA profile, even with DNA amounts lower than 0.5 ng. In the presence of inhibitors, the IAC₉₀ signal was still present after all human DNA loci fail to amplify; in contrast, the IAC₄₁₀ signal was reduced or absent at low levels of inhibition. Amplification of the two IACs provided an internal PCR control and allowed partial profiles caused by inhibition to be distinguished from degraded DNA profiles.

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Section: Generation Of the Iacsmentioning

confidence: 99%

Development of internal amplification controls for DNA profiling with the AmpFℓSTR^® SGM Plus^® kit

et al. 2011

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“…It may be possible to use alternative light sources to detect such stains [26,27], but that was not investigated in this study. Haem from blood and indigo blue dye from denim can have an inhibitory effect on PCR [24,[28][29][30]. Little or no inhibition was observed in this study with streptococcal DNA being detected on denim even after washing.…”

Section: Discussionmentioning

confidence: 67%

Using oral microbial DNA analysis to identify expirated bloodspatter

et al. 2010

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Distinguishing expirated bloodstains (blood forced by airflow out of the nose, mouth or a chest wound) from impact spatter (blood from gunshots, explosives, blunt force trauma and/or machinery accidents) is an important challenge in forensic science. Streptococcal bacteria are only found in the human mouth and saliva. This study developed a polymerase chain reaction (PCR) method that detects DNA from these bacteria as a sensitive tool to detect the presence of saliva. The PCR method was very specific to human oral streptococci, with no PCR product being made from human DNA or DNA from other microbes that were tested. It was also very sensitive, detecting as little as 60 fg of target DNA. The PCR amplification gave product with 99 out of 100 saliva samples tested. PCR was not inhibited by the presence of blood and could detect target DNA in expirated bloodstains in a range of materials and for up to 92 days after deposit on cardboard or cotton fabric. In a blind trial, the PCR method was able to distinguish three mock forensic samples that contained expirated blood from four that did not. Our data show that bacteria present in the oral cavity can be detected in bloodstains that contain saliva and therefore can potentially be used as a marker in forensic work to distinguish mouth-expirated bloodstains from other types of bloodstains.

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“…This loss may be owing to the picric acid component as highly acidic solutions will depurinate nucleic acids (18) damaging DNA. In addition, indigo carmine, used in the textile industry for dyeing denim, is a known inhibitor of PCR (19), further causing low yields. The use of nuclear fast red stained paraffin‐embedded tissues before LMD has produced a superior yield over other histological stains (17) and, used alone, may be an approach to increasing yield if found sufficient for morphological identification.…”

Section: Discussionmentioning

confidence: 99%

Laser Microdissection Separation of Pure Spermatozoa from Epithelial Cells for Short Tandem Repeat Analysis^*

Sanders

Sanchez

Ballantyne

et al. 2006

Journal of Forensic Sciences

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Short tandem repeat (STR) analysis is a valuable tool in identifying the source of biological stains, particularly from the investigation of sexual assault crimes. Difficulties in analysis arise primarily in the interpretation of mixed genotypes when cell separation of the sexual assailant's sperm from the victim's cells is incomplete. The forensic community continues to seek improvements in cell separation methods from mixtures for DNA typing. The feasibility of applying laser microdissection (LMD) technology to precisely separate sexual assault cell mixtures by visual inspection coupled with laser dissection was assessed through three experiments. First, various histological stains were evaluated for use with LMD and DNA analysis. Second, different DNA isolation methods were evaluated on LMD-collected cells. Finally, STR analysis was performed on LMD-separated sperm cells from mixtures of semen and female buccal epithelial cells. The results indicated (a) hematoxylin/eosin staining performed best in its ability to differentiate sperm and epithelial cells while exhibiting the least negative effect on further downstream analysis; (b) both QIAamp and Lyse-N-Go methods were useful for recovery of DNA from LMD-collected sperm cells; and (c) LMD separation provided clear STR profiles of the male donor with the absence of any additional alleles from the female donor. This report describes an efficient, low-manipulation LMD method for the efficient separation of spermatozoa from two-donor sperm/epithelial cell mixtures.

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An improved method for STR analysis of bloodstained denim

Cited by 39 publications

References 6 publications

Development of internal amplification controls for DNA profiling with the AmpFℓSTR^® SGM Plus^® kit

Development of internal amplification controls for DNA profiling with the AmpFℓSTR^® SGM Plus^® kit

Using oral microbial DNA analysis to identify expirated bloodspatter

Laser Microdissection Separation of Pure Spermatozoa from Epithelial Cells for Short Tandem Repeat Analysis^*

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An improved method for STR analysis of bloodstained denim

Cited by 39 publications

References 6 publications

Development of internal amplification controls for DNA profiling with the AmpFℓSTR® SGM Plus® kit

Development of internal amplification controls for DNA profiling with the AmpFℓSTR® SGM Plus® kit

Using oral microbial DNA analysis to identify expirated bloodspatter

Laser Microdissection Separation of Pure Spermatozoa from Epithelial Cells for Short Tandem Repeat Analysis*

Contact Info

Product

Resources

About

Development of internal amplification controls for DNA profiling with the AmpFℓSTR^® SGM Plus^® kit

Development of internal amplification controls for DNA profiling with the AmpFℓSTR^® SGM Plus^® kit

Laser Microdissection Separation of Pure Spermatozoa from Epithelial Cells for Short Tandem Repeat Analysis^*