Allele Peak Heights Research Articles

Half-volume validation of the NGM Detect™ PCR Amplification Kit and its application on degraded casework samples.

The NGM Detect™ PCR Amplification Kit was designed particularly for genotyping degraded casework samples. This study aimed to validate the half-volume amplification of the kit and to present its successful long-term application. The validation was performed in accordance to the corresponding guidelines of the Scientific Working Group on DNA analysis methods and the European Network of Forensic Science Institutes. For validation parameters, such as sensitivity, reproducibility, and repeatability, polymerase chain reactions (PCR) were set up both manually and robotically, applying 29 cycles. For PCRs with sub-optimal DNA input (≤0.5 ng) the cycle numbers were increased to 31. Regardless of the PCR preparation method, the optimal 0.5 ng DNA input produced optimal allelic peak heights with no allelic dropout. The first alleles that failed to amplify started to appear at the level of 0.0375 ng input DNA, although the manually prepared PCRs produced fewer missing alleles. In this case, the raised cycle number produced 1.9% and 4.4% of dropout for manually and for robotically set up PCRs, respectively. In the case of 84 degraded casework samples, PCRs were prepared only by hand. The kit was able to provide informative profiles for 78.57%, 70.37%, and 69.77% for lowly, moderately, and highly degraded samples, respectively. Allelic dropouts were 26.05%, 44.88%, and 51.23% for the same groups. According to our results, we strongly recommend using the NGM Detect™ Kit in half-volume PCR system and encourage the usage of the kit in the particular cases when other kits fail to produce a complete DNA profile.

Assessing DNA Degradation through Differential Amplification Efficiency of Total Human and Human Male DNA in a Forensic qPCR Assay.

The assessment of degradation is crucial for the analysis of human DNA samples isolated from forensic specimens. Forensic quantitative PCR (qPCR) assays can include multiple targets of varying amplicon size that display differential amplification efficiency, and thus different concentrations, in the presence of degradation. The possibility of deriving information on DNA degradation was evaluated in a forensic qPCR assay not specifically designed to detect DNA fragmentation, the Plexor HY (Promega), by calculating the ratio between the estimated concentrations of autosomal (99 bp) and Y-chromosomal (133 bp) targets ("[Auto]/[Y]"). The [Auto]/[Y] ratio measured in 57 formalin-fixed, paraffin-embedded samples was compared to a quality score (QS) calculated for corresponding STR profiles using quantitative data (allele peak height). A statistically significant inverse correlation was observed between [Auto]/[Y] and QS (R = -0.65, p < 0.001). The [Auto]/[Y] values were highly correlated (R = 0.75, p < 0.001) with the "[Auto]/[D]" values obtained using the PowerQuant (Promega) assay, expressly designed to detect DNA degradation through simultaneous quantification of a short (Auto) and a long (D) autosomal target. These results indicate that it is possible to estimate DNA degradation in male samples through Plexor HY data and suggest an alternative strategy for laboratories lacking the equipment required for the assessment of DNA integrity through dedicated qPCR assays.

A fast and inexpensive genotyping system for the simultaneous analysis of human and Aedes albopictus short tandem repeats

BackgroundDetermination of the interactions between hematophagous mosquitoes and their human hosts is of great importance for better understanding the transmission dynamics of mosquito-borne arboviruses and developing effective strategies to mitigate risk. Genetic analysis of human and mosquito DNA can play a key role in this, but commercial kits for human short tandem repeat (STR) genotyping are expensive and do not allow for the simultaneous STR analysis of host and vector DNA. Here, we present an inexpensive and straightforward STR-loci multiplex system capable of simultaneously amplifying Aedes albopictus and human STRs from blood-fed mosquitoes. Additionally, we examine the effect of storage methods and post-feeding time on the integrity of host DNA.MethodsThirty-five STRs (16 human and 19 Ae. albopictus STRs) subdivided in three multiplexes were tested for amplification and scoring reliability. Under laboratory conditions we compared the efficacy of two preservation methods (absolute ethanol vs lysis buffer) on the integrity of host DNA in Ae. albopictus blood meals. We also evaluated the effect of post-feeding time by sacrificing blood-fed mosquitoes at different time intervals after feeding, and we assessed our ability to detect multiple feedings. To determine if the system can be employed successfully under field conditions, we carried out a preliminary study using field-collected Ae. albopictus.ResultsAll 35 STRs amplified consistently in the laboratory. Lysis buffer performed better than absolute ethanol in terms of allele peak height and clarity of electropherograms. Complete human DNA profiles could be obtained up to 48 h following the blood meal. Analysis of multiple feedings confirmed that peak heights can be used as a proxy to determine post-feeding time and thus derive the number of different people bitten by a mosquito. In the field trial, amplification was successful for 32 STRs. We found human DNA signal in 38 of the 61 field-collected mosquitoes (62%), of which 34 (89%) had ingested a single blood meal, while four (11%) contained double meals.ConclusionsOur new genotyping system allows fast and reliable screening of both host and vector species, and can be further adapted to other mosquito species living in close contact with humans.Graphical

Evaluation of large-scale highly polymorphic microhaplotypes in complex DNA mixtures analysis using RMNE method

DNA mixture interpretation is one of the most challenging problems in forensics. Complex DNA mixtures are more difficult to analyze when there are more than two contributors or related contributors. Microhaplotypes (MHs) are polymorphic genetic markers recently discovered and employed in DNA mixture analysis. However, the evidentiary interpretation of the MH genotyping data needs more debate. The Random Man Not Excluded (RMNE) method analyzes DNA mixtures without using allelic peak height data or the number of contributors (NoC) assumptions. This study aimed to assess how well RMNE interpreted mixed MH genotyping data. We classified the MH loci from the 1000 Genomes Project database into groups based on their Ae values. Then we performed simulations of DNA mixtures with 2–10 unrelated contributors and DNA mixtures with a pair of sibling contributors. For each simulated DNA mixture, incorrectly included ratios were estimated for three types of non-contributors: random men, parents of contributors, and siblings of contributors. Meanwhile, RMNE probability was calculated for contributors and three types of non-contributors, allowing loci mismatch. The results showed that the MH number, the MH Ae values, and the NoC affected the RMNE probability of the mixture and the incorrectly included ratio of non-contributors. When there were more MHs, MHs with higher Ae values, and a mixture with less NoC, the RMNE probability, and the incorrectly included ratio decreased. The existence of kinship in mixtures complicated the mixture interpretation. Contributors’ relatives as non-contributors and related contributors in the mixture increased the demands on the genetic markers to identify the contributors correctly. When 500 highly polymorphic MHs with Ae values higher than 5 were used, the four individual types could be distinguished according to the RMNE probabilities. This study reveals the promising potential of MH as a genetic marker for mixed DNA interpretation and the broadening of RMNE as a parameter indicating the relationship of a specific individual with a DNA mixture in the DNA database search.

Genetic Polymorphism of 38 Y-chromosome Short Tandem Repeats in Beijing Han Population from China

Abstract Objective: To investigate 38 Y-chromosome short tandem repeat (Y-STR) genetic polymorphisms in Beijing Han and analyze the genetic distance with neighboring or linguistically similar populations. Materials and Methods: In the study, we selected 531 unrelated male individuals of Beijing Han, and the results were statistically analyzed by testing with GSTAR™ 41Y reagents. Results: The allele peak heights were balanced among the Y loci, the amplified fragment ranged from 100 to 500 bps. A total of 531 haplotypes were detected in 531 samples. Eight null genotypes were observed on locus DYS448. One and three double alleles were observed on single-copy locus DYS576 and DYS19, respectively. DYS385 a/b, DYF387S1 a/b, and DYS527 a/b were more common in double copies, but 3, 13, and 11 triple alleles were detected, respectively. The gene diversity values of Y-STRs except DYS391, DYS438, and DYS645 were &gt;0.5. Twenty-seven Y-STRs of Beijing Han population were selected for genetic distance comparison with 17 populations including Changchun Han, with Rst values ranging from 0.0002 to 0.1703. Conclusion: The 38 Y-STRs in this study have strong male lineage identification ability and have great potential for individual identification, kinship identification, Y-STR database construction, and genetic relationship research.

Internal Validation of MaSTR™ Probabilistic Genotyping Software for the Interpretation of 2-5 Person Mixed DNA Profiles.

Mixed human deoxyribonucleic acid (DNA) samples present one of the most challenging pieces of evidence that a forensic analyst can encounter. When multiple contributors, stochastic amplification, and allele drop-out further complicate the mixture profile, interpretation by hand becomes unreliable and statistical analysis problematic. Probabilistic genotyping software has provided a tool to address complex mixture interpretation and provide likelihood ratios for defined sets of propositions. The MaSTR™ software is a fully continuous probabilistic system that considers a wide range of STR profile data to provide likelihood ratios on DNA mixtures. Mixtures with two to five contributors and a range of component ratios and allele peak heights were created to test the validity of MaSTR™ with data similar to real casework. Over 280 different mixed DNA profiles were used to perform more than 2600 analyses using different sets of propositions and numbers of contributors. The results of the analyses demonstrated that MaSTR™ provided accurate and precise statistical data on DNA mixtures with up to five contributors, including minor contributors with stochastic amplification effects. Tests for both Type I and Type II errors were performed. The findings in this study support that MaSTR™ is a robust tool that meets the current standards for probabilistic genotyping.

A new implementation of a semi-continuous method for DNA mixture interpretation

A new calculation module within the PopStats module of the CODIS software package, based on the underlying mathematics presented in the MixKin software package, has been developed for assigning the Likelihood Ratio (LR) of DNA mixture profiles. This module uses a semi-continuous model that allows for population structure and allelic drop-out and drop-in but does not require allelic peak heights or other laboratory-specific parameters. This new implementation (named SC Mixture), like MixKin, does not specify or estimate a probability of drop-out. Instead, each contributor to a mixture has an independent drop-out rate, and the probability of the mixture profile for a specified proposition concerning the contributors is integrated over the range of possible drop-out rates. The allelic drop-in rate and the population structure parameter, theta, used by the software are specified by the user. The user can examine up to five contributors to a mixture, however, conditioning on assumed contributors and limiting the number of unknowns in both numerator and denominator hypotheses greatly improves performance. We report results from an extensive validation study performed for ten mixtures with each of one (single source), two, three, four, or five contributors, with four combinations of drop-in rate and a population structure parameter. Each mixture was run as a complete profile or with the random removal of alleles to simulate drop-out. All 1620 combinations were evaluated with PopStats, MixKin, and LRmix and considerable consistency was found among the results with all three packages.

High-quality data from a forensically relevant single-cell pipeline enabled by low PBS and proteinase K concentrations.

Interpreting forensic DNA signal is arduous since the total intensity is a cacophony of signal from noise, artifact, and allele from an unknown number of contributors (NOC). An alternate to traditional bulk-processing pipelines is a single-cell one, where the sample is collected, and each cell is sequestered resulting in n single-source, single-cell EPGs (scEPG) that must be interpreted using applicable strategies. As with all forensic DNA interpretation strategies, high quality electropherograms are required; thus, to enhance the credibility of single-cell forensics, it is necessary to produce an efficient direct-to-PCR treatment that is compatible with prevailing downstream laboratory processes. We incorporated the semi-automated micro-fluidic DEPArray™ technology into the single-cell laboratory and optimized its implementation by testing the effects of four laboratory treatments on single-cell profiles. We focused on testing effects of phosphate buffer saline (PBS) since it is an important reagent that mitigates cell rupture but is also a PCR inhibitor. Specifically, we explored the effect of decreasing PBS concentrations on five electropherogram-quality metrics from 241 leukocytes: profile drop-out, allele drop-out, allele peak heights, peak height ratios, and scEPG sloping. In an effort to improve reagent use, we also assessed two concentrations of proteinase K. The results indicate that decreasing PBS concentrations to 0.5X or 0.25X improves scEPG quality, while modest modifications to proteinase K concentrations did not significantly impact it. We, therefore, conclude that a lower than recommended proteinase K concentration coupled with a lower than recommended PBS concentration results in enhanced scEPGs within the semi-automated single-cell pipeline.

Direct STR typing from human bones

Identification by STR analysis of bones is time-consuming, mainly due to the lengthy decalcification required and complex DNA extraction process. To streamline this process, we developed a direct STR typing protocol from bone samples. We optimized bone sample amounts using femur and tibia and two commercial PCR kits (Identifiler™ Plus and IDplex Plus kits). Optimally, 100 mg of bone powder in 300 µL PBS buffer was heated at 98 °C for three minutes to produce a supernatant for DNA amplification. IDplex Plus performed better than Identifiler™ Plus in terms of allele recovery and peak height. Fifteen samples of each of seven bone elements (1st distal phalange of hand, capitate, femur, metacarpal 4, patella, talus, and tibia; N = 105) were then subjected to direct STR typing with the optimized protocol, and 94.3% were high partial to full profiles. The performance of the developed protocol was similar for all bone elements. Median peak heights were significantly better in profiles of cancellous bone than compact bone (p = 0.033) and significantly different across the bone elements (p < 0.001). Ten casework samples from various conditions and up to 7-year-PMI were subjected to both direct STR and conventional STR typing. No significant difference in the number of alleles was seen (95% HDI of −13.5 to 5.15). As well as being rapid, convenient, and safe, the protocol could help improve STR typing from bones.

A proof-of-principle study on implementing polymerase chain displacement reaction (PCDR) to improve forensic low-template DNA analysis

Polymerase chain reaction (PCR) plays an important role in forensic DNA analysis. However, the amplification of low-template DNA (LTDNA) samples usually encounters unsatisfactory results for the limited efficiency of PCR, which would interfere with the subsequent profile interpretation. Polymerase chain displacement reaction (PCDR) is a highly-efficient technique characterized by combining PCR and strand displacement reaction into a single PCDR cycle. This study explored the feasibility of PCDR for improving forensic LTDNA analysis. STR markers commonly used in forensic genetics were subjected to PCDR amplification and capillary electrophoresis detection. The results of singleplex reactions indicated that PCDR surpassed original PCR in efficiency for STR amplification. The average peak height of alleles in PCDR profiles was linearly correlated to the number of outer primers adopted for initiating the strand displacement process. Further, we assessed the multiplexing potential of PCDR by incorporating 17 STRs included in the expanded CODIS core loci and Amelogenin gene into a multiplex PCDR system. For pristine DNA templates ranged from 200 pg to 12.5 pg, the multiplex PCDR system consistently exhibited higher allele peak height as well as less allele dropout compared to the multiplex PCR references. Meanwhile, a significant reduction of stutter ratio was extensively observed in PCDR profiles. We also tested mock casework samples to verify the practical ability of multiplex PCDR for LTDNA detection. With DNA input varying from 48.1 pg to 6.6 pg, the multiplex PCDR system consistently obtained more allelic information than multiplex PCR methods. Our data collectively suggested that it is feasible to apply PCDR in forensic LTDNA analysis.

Calcium-Alginate Tissue Gels (CATG): Proof-of-concept biomaterial development

Hydrogels are desirable materials to the field of forensic science and offer many advantages for use as tissue simulants in research and training scenarios. In this work, we demonstrate a proof-of-concept study for our biomaterial described as the Calcium-Alginate Tissue Gel (CATG). CATG biomaterials integrate functional DNA strands designed to amplify with known human primer sets for genetic profiling. Our range of CATG materials demonstrate successful DNA extraction, PCR amplification and genotyping when both fresh and aged for 21 days. The rheological properties of the CATGs were measured and the incorporation of DNA into the CATGs was assessed. Overall, the CATGs demonstrated increased viscoelastic behavior with the addition of DNA. In addition, two methods of sampling were considered, where it was found that cutting a sample of the dried CATG produced higher allele peak heights in the genotype compared to swabbing. Overall, our CATG biomaterials can be designed for multiple applications in forensic science with tunable properties for various training and research needs.

Modeling allelic analyte signals for aSTRs in NGS DNA profiles.

We describe an adaption of Bright et al.'s work modeling peak height variability in CE-DNA profiles to the modeling of allelic aSTR (autosomal short tandem repeats) read counts from NGS-DNA profiles, specifically for profiles generated from the ForenSeq™ DNA Signature Prep Kit, DNA Primer Mix B. Bright et al.'s model consists of three key components within the estimation of total allelic product-template, locus-specific amplification efficiencies, and degradation. In this work, we investigated the two mass parameters-template and locus-specific amplification efficiencies-and used MLE (maximum likelihood estimation) and MCMC (Markov chain Monte Carlo) methods to obtain point estimates to calculate the total allelic product. The expected read counts for alleles were then calculated after proportioning some of the expected stutter product from the total allelic product. Due to preferential amplicon selection introduced by the sample purification beads, degradation is difficult to model from the aSTR outputs alone. Improved modeling of the locus-specific amplification efficiencies may mask the effects of degradation. Whilst this model could be improved by introducing locus specific variances in addition to locus specific priors, our results demonstrate the suitability of adapting Bright et al.'s allele peak height model for NGS-DNA profiles. This model could be incorporated into continuous probabilistic interpretation approaches for mixed DNA profiles.

Bayesian regression analysis of stutter in DNA mixtures

Probabilistic genotyping methods use a hierarchical probability model in deconvolution of DNA mixtures. The parameters of the model, including the stutter which are required to calculate the expected values of peak heights, are estimated in the validation process. Linear modeling of stutter, as a common artifact in DNA genotyping, has been reported previously. The typically right-skewed error distribution and non-negativeness of stutter to its allele peak heights ratios make generalized linear models preferable, especially Bayesian analogs, which allow even more flexibility. In this paper, we show how such models can be fitted and applied with the aid of Markov chain Monte Carlo methods.

Single step strategy for estimating unknown STR allele frequencies in a population

Constant growth of forensic DNA databases requires new STR markers to be developed, tested and validated. Aiming to simplify the process of obtaining allele frequency distributions of new STR loci included in new megaplexes, a single step protocol was developed. This goal was achieved by pooling samples of genetically unrelated donors, previously genotyped (n = 99), containing the same amount of DNA (total DNA = 4.5 ng/ul) as measured by fluorometry. Suitability of the method was tested by amplifying commercial kits: GlobalFiler (GF), Verifiler Express (VFE) and Y-filer plus (YFP). Allele peak heights was used to estimate allele frequency and compared with the actual frequency calculated by counting method of the individual pooled samples. The multinomial goodness of fit test was performed for testing statistical differences.In case of VFE, 4.5 ng was the DNA amount that could best estimate the allele frequency distribution; 1 ng for GF and YFP. For simple, complex and compound makers, the highest allele frequencies were correctly estimated (p > 0.05) but in some cases, it was not able to detect alleles in very low frequencies, 2/198 or below, either with VFE or GF.In case of D12S391, either for VFE and GF, some microvariants with frequency of 2, 3 or 5/198 were not assigned although they were present in the profiles, probably due to POP7 resolution. In case of YFP, some alleles were not detected at DYS385.Nevertheless, with some limitations, the proposed strategy proved to be fast and efficient for major allele frequency estimation. This simple approach generates an empirical distribution that allows for the estimation of population allele frequencies in a single step assay.

Automated estimation of the number of contributors in autosomal short tandem repeat profiles using a machine learning approach

The number of contributors (NOC) to (complex) autosomal STR profiles cannot be determined with absolute certainty due to complicating factors such as allele sharing and allelic drop-out. The precision of NOC estimations can be improved by increasing the number of (highly polymorphic) markers, the use of massively parallel sequencing instead of capillary electrophoresis, and/or using more profile information than only the allele counts.In this study, we focussed on machine learning approaches in order to make maximum use of the profile information. To this end, a set of 590 PowerPlex® Fusion 6C profiles with one up to five contributors were generated from a total of 1174 different donors. This set varied for the template amount of DNA, mixture proportion, levels of allele sharing, allelic drop-out and degradation. The dataset contained labels with known NOC and was split into a training, test and hold-out set. The training set was used to optimize ten different algorithms with selection of profile characteristics. Per profile, over 250 characteristics, denoted ‘features’, were calculated. These features were based on allele counts, peak heights and allele frequencies. The features that were most related to the NOC were selected based on partial correlation using the training set. Next, the performance of each model (=combination of features plus algorithm) was examined using the test set. A random forest classifier with 19 features, denoted the ‘RFC19-model’ showed best performance and was selected for further validation. Results showed improved accuracy compared to the conventional maximum allele count approach and an in-house nC-tool based on the total allele count. The method is extremely fast and regarded useful for application in forensic casework.

Computation of marginal distributions of peak-heights in electropherograms for analysing single source and mixture STR DNA samples

Current models for single source and mixture samples, and probabilistic genotyping software based on them used for analysing STR electropherogram data, assume simple probability distributions, such as the gamma distribution, to model the allelic peak height variability given the initial amount of DNA prior to PCR amplification. Here we illustrate how amplicon number distributions, for a model of the process of sample DNA collection and PCR amplification, may be efficiently computed by evaluating probability generating functions using discrete Fourier transforms.

A novel FTA™ elute card collection method that improves direct DNA amplification from bloodstained concrete

Concrete is a common construction material found in residential and commercial buildings, bridges and parking lots that is a composite matrix containing aggregate held together with cement. The porous nature of concrete can make the collection and genotyping of biological fluids, such as blood, challenging. Forensic evidence can become embedded within the matrix, potentially reducing the amount of DNA available for analysis. In forensic science, “direct” amplification refers to a genotyping method that amplifies a DNA profile directly from a sample without DNA extraction, saving time and money. We investigated a novel application of Whatman™ FTA™ Elute cards in their ability to directly amplify PowerPlex® Fusion and Y23 profiles from minute amounts of blood that had been deposited on different concrete structures. In comparison to traditional collection methods, directly profiling blood stained construction materials using FTA™ Elute cards increased the percentage loci amplified and significantly improved both allele peak height and peak height ratio while reducing allelic drop-out. FTA™ Elute cards can provide a reliable, inexpensive and superior alternative to traditional methods.

Low-template DNA: A single DNA analysis or two replicates?

This study investigates the following two questions: (1) Should the DNA analyst concentrate the DNA extract into a single amplification or should he/she split it up to do two replicates? (2) Given the electropherogram obtained from a first analysis, is it worthwhile for the DNA analyst to invest in obtaining a second replicate? A decision-theoretic approach addresses these questions by quantitatively expressing the expected net gain (ENG) of each DNA analysis of interest. The results indicate that two replicates generally have a greater ENG than a single DNA analysis for DNA quantities capable of producing two replicates having an average allelic peak height as low as 43rfu. This supports the position that two replicates increase the information content with regard to a single analysis.

New stutter ratio distribution for DNA mixture interpretation based on a continuous model.

In forensic science, DNA mixture interpretation is traditionally based on a binary model, which does not account for peak-height information in DNA profiles. In recent years, some countries have adopted a continuous model in which peak heights are used and stochastic effects are considered to enable rigorous calculation of likelihood ratios. However, this model requires certain biological parameters which affect the expected allelic and stutter peak heights. In this paper, we focused on estimating the distribution of the stutter ratio (SR) in 15 short tandem repeat loci in relation to the allele repeat number. We estimated the SR values of 234 single-source DNA samples by using a commercially available kit. In all loci except for D8S1179, D21S11, and D2S1338, a simple log-normal distribution model was fitted to the variability of SR. For D21S11, we developed a new distribution model in which distinct log-normal distributions between complete and incomplete repeat units are used (a separate log-normal distribution model). For D8S1179 and D2S1338, we developed another new distribution model that mixes two log-normal distributions to explain two types of repeat structures appearing within the same number of allele repeats. These two models were fitted to the observed SR values more accurately than the simple log-normal distribution model. We expected these new SR models to be applied to DNA mixture interpretation based on a continuous model.

Allele Peak Height Imbalance Ratios between Different Commercially Available STR Kits on Sexual Violence Crime Scene Evidences

STR 분석기술은 법과학 분야를 포함한 다양한 분야에서 폭넓게 활용되고 있는 가장 효과적이고 안정적인 방법이다. 그러나 심하게 손상된 시료나 미량 시료의 경우, 증폭과정에서 나타나는 여러 가지 인공 산물들로 인하여 유효성 있는 결과를 확보하지 못하는 경우가 종종 발생하게 된다. 특히, 대립유전 자간 불균형의 경우, 혼합반 시료의 분석에 있어서 결과 추정 시 데이터 해석의 혼란을 야기하는 만큼 그에 대한 연구의 필요성이 제기되고 있다. 본 연구에서는 성범죄 현장시료를 대상으로 비혼합반 시료 461종을 선별한 후, 3가지 상용화 STR kit를 이용하여 대립유전자간 불균형 비율을 분석하였다. 그 결과 대립유전자간 불균형 비율이 상대적으로 높은 5개 좌위(Amelogenin, D13S317, D18S51, FGA 및 Penta E)와 상대적으로 낮은 3개 좌위(TH01, TPOX 및 D3S1358)를 선별할 수 있었다. 이러한 결과를 바탕으 로 데이터 분석에 있어서 대립유전자간의 불균형 비율을 감안하여 분석한다면, 결과 분석 시 범할 수 있 는 해석의 오류를 최소화 시키는데 도움이 될 것으로 판단된다.Although STR typing is the most representative methods in the forensic field, it has a limits such as stutter, allele drop-out, non-template addition, peak height imbalance in the damaged DNA or a small amount of crime scene evidence. Specially, peak height imbalance is one of the important factors that could cause the debate to allow researchers to interpret the DNA genotyping results. In this study, we selected the 461 samples of sexual violence crime scene evidence in order to determine the peak height imbalance ratios of the three types of commercially available kits (AmpFlSTR？？ Identifiler Plus Kit, PowerPlex？？ 16 HS System Kit, PowerPlex？？ Fusion System Kit) which were currently used for STR analysis. As a result, we were able to determine the high level group (Amelogenin, D13S317, D18S51, FGA, Penta E) which was required a careful analysis of the STR results because of the peak height imbalance ratios high. In addition, we have confirmed the low level group (TH01, TPOX, D3S1358) which a stable ratios of peak height imbalance. These results suggest that may help to minimize the STR data analysis error for mixture samples of crime scene evidence.