Search

Microbiome studies have contributed to many fields, such as healthcare and medicine; however, these studies are relatively limited in forensics. Microbiome analyses can provide information, such as geolocation and ancestry information, when short tandem repeat (STR) profiling fails. In this study, methods for DNA extraction and sampling from the skin and saliva were optimized for the construction of a Korean Forensic Microbiome Database (KFMD). DNA yields were estimated using four DNA extraction kits, including two automated kits (Maxwell® FSC DNA IQ™ Casework Kit and PrepFiler™ Forensic DNA Extraction Kit, updated) and two manual kits (QIAamp DNA Mini Kit and QIAamp DNA Micro Kit) commonly used in forensic DNA profiling laboratories. Next-generation sequencing of the 16S rRNA V4 region was performed to analyze microbial communities in samples. The Bacterial Transport Swab with Liquid Media (NobleBio), two cotton swabs (PoongSung and Puritan), and nylon-flocked swabs (NobleBio and COPAN) were tested for DNA recovery. The PrepFiler and Maxwell kits showed the highest yields of 3.884 ng/μL and 23.767 ng/μL from the scalp and saliva, respectively. With respect to DNA recovery, nylon-flocked swabs performed better than cotton swabs. The relative abundances of taxa sorted by DNA extraction kits were similar contributions; however, with significant differences in community composition between scalp and saliva samples. Lawsonella and Veillonella were the most abundant genera in the two sample types. Thus, the Maxwell® FSC DNA IQ™ Casework Kit and nylon-flocked swab (NobleBio) were optimal for DNA extraction and collection in microbiome analyses.

The InnoTyper 21® Human Identification kit consists of amelogenin and 20 bi-allelic Alus, retrotransposon markers existing abundantly in human genome. The InnoTyper 21® kit produces shorter amplicons (60-125 bp) than conventional short tandem repeat (STR) genotyping kit, then it is effective on the analysis of challengeable forensic samples including insufficient or highly degraded DNAs. Also, as the genotyping with InnoTyper21® kit is compatible with PCR and capillary electrophoresis, it is easy to incorporate into the workflow in forensic laboratories. In the internal validation of InnoTyper21® kit on sensitivity, degradation, and mixture studies for the evaluation in this study, we acquired full profiles on analyzing small concentration DNA (as low as 25 pg) and highly degraded DNA (up to 105 degradation index value). Through the Korean population study, forensic statistical parameters were investigated and a specific variant of T insertion in NBC51 was confirmed in six samples. Comparison of Korean population with five populations or 1000 Genomes Project data show Korean specific substructure. It is expected that the InnoTyper 21® kit will be used into the actual forensic cases, utilizing the population study investigated through this research.

The recent studies reported that DNA methylation markers show changes with age, and expected that the DNA methylation markers can be effectively used for estimation of age in forensic genetics. In this study, we applied droplet digital PCR (ddPCR) method to investigate the DNA methylation pattern in the CpG sites, and we constructed an age prediction model based on the ddPCR method. The ddPCR is capable of highly sensitive quantitation of nucleic acid and detection of sequence variations in gene by separating the sample into large number of partitions and clonally amplifying nucleic acids in each partition. We extracted DNA from saliva samples collected from several age groups. The DNA was bisulfite converted and subjected to ddPCR using specifically designed primers and probes. The methylation ratio of each sample was calculated and correlation between the methylation ratio and the chronological age was analyzed. In the results, methylated DNA ratio at the 4 CpG sites (cg14361627, cg14361627, cg08928145 and cg07547549) showed strong correlation with chronological age. Percent-methylation values at 4 CpG markers and chronological ages of the 76 individuals were analyzed by multiple regression analysis, and we constructed an age prediction model. We observed a strong correlation (Spearman’s rho = 0.922) between predicted and chronological ages of 76 individuals with a MAD from chronological age of 3.3 years. Collectively, the result in this study showed the potential applicability of ddPCR to predict age from saliva.

The analysis of Y-SNPs from crime scene samples is helpful for investigators in narrowing down suspects by predicting biogeographical ancestry. In this study, a PCR-reverse blot hybridization assay (REBA) for predicting Y-chromosome haplogroups was employed to determine the major haplogroups worldwide, including AB, DE, C, C3, F, K, NO, O, O2, and O3 and evaluated. The REBA detects nine biallelic Y chromosome markers (M9, M89, M122, M145, M175, M214, M217, P31, and RPS4Y711) simultaneously using multiple probes. The REBA for Y-single nucleotide polymorphisms (SNP) genotyping was performed using 40 DNA samples from Asians—14 Koreans, 10 Indonesians, six Chineses, six Thais, and four Mongolians. 40 Asian samples were identified as haplogroup O2 (40%), O3 (32.5%), C3 (17.5%), O (7.5%) and K (2.5%). These cases were confirmed by DNA sequence analysis (κ = 1.00; P

This study developed a new method for forensic saliva identification using three oral bacteria, Streptococcus salivarius, Streptococcus sanguinis, and Neisseria subflava, combined with a real-time polymerase chain reaction (RT-PCR) system we called OB mRT-PCR. Analytical sensitivity results showed that the target bacteria were amplified at 102–107 copies/reaction, and analytical specificity was assessed using 24 other viruses, bacteria, and protozoa. To evaluate the OB mRT-PCR kit for forensic applications, saliva from 140 Korean individuals was tested, and at least two target bacteria were detected in all the samples. Additional studies on non-saliva samples demonstrated the specificity of the kit. Comparison of the kit with two conventional saliva test methods, the SALIgAE and RSID-Saliva assays, indicated that it was more sensitive and applicable to saliva samples in long-term storage (up to 14 weeks). Additionally, through amplification of mock forensic items and old DNA samples (isolated without lysis of the bacterial cells, regardless of their Gram-positivity), we found that the kit was applicable to not only saliva swabs, but also DNA samples. We suggest that this simple RT-PCR-based experimental method is feasible for rapid on-site analysis, and we expect this kit to be useful for saliva detection in old forensic DNA samples.

In this paper, we describe the development of a novel method to detect oral bacteria by combining direct polymerase chain reaction (direct PCR) with an immunochromatographic strip (ICS), enabling the identification of saliva in forensic samples. Direct PCR was first used to directly amplify specific oral bacterial sequences (from Streptococcus sanguinis and Streptococcus salivarius) from swab samples, circumventing the need for tedious sample preparation steps such as cell lysis and DNA extraction and purification. The resultant amplicons were then colorimetrically detected on an ICS, a much more convenient, cost-effective, and user-friendly detection method than those currently available, thereby allowing the presence or absence of the target oral bacteria to be determined with the naked eye. Moreover, the entire analysis process was performed rapidly and with ease using this combination of direct PCR amplification from swab samples and ICS-based amplicon detection. This method successfully detected S. sanguinis and S. salivarius in most of the saliva swab samples tested, and returned negative results using blood, semen, urine, and vaginal fluid swab samples. Furthermore, S. sanguinis and S. salivarius were detected in a large number of mock forensic samples using this technique, which suggests that direct PCR and ICS-based detection of oral bacteria is sufficient to demonstrate the presence of saliva. Thus, we believe that the proposed method could be very useful for the identification of saliva in forensic applications.

To increase the discrimination power of human identification in forensic and to evaluate expanded short tandem repeat (STR) loci, we examined a Korean sample population of 2000 unrelated individuals using both GlobalFiler™ PCR Amplification kit and PowerPlex® Fusion System. Allele frequencies for the 20 STR loci (D3S1358, vWA, D16S539, CSF1PO, TPOX, D8S1179, D21S11, D18S51, D2S441, D19S433, TH01, FGA, D22S1045, D5S818, D13S317, D7S820, D10S1248, D1S1656, D12S391, and D2S1338) were calculated. Allele frequencies of the 20 STR loci would be useful for personal identification and paternity testing in forensics and as a standard reference database in Korea.

Recently, a method of identifying body fluids using DNA methylation has been developed (Frumkin et al., 2011). An existing multiplex assay using 9 CpG markers could differentiate 5 body fluids: semen, blood, saliva, menstrual blood, and vaginal fluid. To validate this technique, we evaluated the previously described body fluid identification method by means of single base extension (SBE). DNA methylation was applied to 22 samples in 18 forensic cases; seven of these were semen, three were blood, eight were saliva, three were vaginal fluid, and one was menstrual blood. Total of 18 samples were tested, the DNA methylation profiles were coincident from preliminary tests (acid phosphatase (AP), leucomalachite green (LMG, Sigma Aldrich, St Louis, MO, USA) and SALIgAE®) except one sample which displayed an all-negative result. After applying the DNA methylation method to forensic samples, we determined that it could be very useful for differentiating vaginal secretions from menstrual blood, for which there is no conventional preliminary testing method.

범죄현장 증거물의 DNA 분석에서 2인 이상의 혼합된 DNA 프로필이 종종 검출된다. 법유전학 분야에서는 이러한 혼합 DNA를 분석하고 해석하려는 연구가 지속적으로 이루어지며 관련된 분석 프로 그램도 꾸준히 개발되고 있다. 2010년 ‘DNA 신원확인정보의 이용 및 보호에 관한 법률’이 시행되면서 우 리나라는 국가 DNA 데이터베이스를 운영하기 시작했다. 그러나 혼합형으로 분석된 DNA 데이터는 시스 템의 부재로 검색 및 관리가 효율적으로 이루어지지 않고 있다. 이에 따라 본 연구에서는 이러한 혼합 DNA를 체계적으로 관리하고 검색할 수 있는 시스템인 Forensic Mixed DNA Database Platform (FMDD) 를 개발하였다. FMDD는 혼합 DNA 프로필과 용의자 대조 DNA 프로필간의 상호 검색이 가능하고, 일치 결과에 대한 random match probability (RMP), combined probability of exclusion/inclusion (CPE/CPI), 그리 고 likelihood ratio (LR)의 통계 결과를 제공한다. FMDD에서 제공되는 다양한 기능은 혼합 DNA 프로필 로 분석된 사건을 해결하는데 도움을 줄 뿐만 아니라 신뢰성 있는 결과를 제공할 수 있게 될 것이다

Analysis of single nucleotide polymorphisms (SNPs) in mitochondrial (mt)DNA hypervariable regions (HV) 1/2 is valuable in forensic investigations. We developed a method for mtDNA screening of the HV1 and HV2 regions by melting curve analysis, using peptide nucleic acid (PNA) probes. This method focuses on melting peak patterns obtained by thermal dissociation of PNA/DNA duplexes in amplified mtDNA products. Five PNA probe sets were designed to detect 25 SNPs in the two HV regions. We also detected non-target SNPs based on unexpected melting temperature (Tm) shifts. In fact, 62 SNPs (42 SNPs in HV1 and 20 in HV2) were identified, including the 25 target SNPs. Using this method, 46 melting peak patterns, including 8 pattern groups, were obtained in 60 unrelated individuals. The peak patterns were compared to 55 haplotypes identified by Sanger sequencing. The results obtained from analysis of target mtDNA SNPs were entirely consistent with those obtained by Sanger sequencing. Screening the HV1 and HV2 regions of mtDNA by this method may help minimize unnecessary recourse to full sequence analysis, allows to rapidly exclude samples that do not match evidence and reference samples, and may reduce turnaround times and analysis costs. Overall, this method may be effective and helpful in forensic investigations.

DNA quantification is an essential step for successful multiplex short tandem repeat (STR) polymerase chain reactions (PCR), which are used for confirming identities using human genomic DNA. The new DNA quantification kit, named the National Forensic Service Quantification (NFSQ) kit, simultaneously provides total human DNA concentration, human male DNA concentration, and a DNA degradation index (DI) using multiplex TaqMan fluorescent probes. The NFSQ was validated according to developmental validation guidelines from the SWGDAM and MIQE. NFSQ detected up to 0.00128 ng/μL and could detect male DNA up to a 1:8000 ratio of male to female DNA. In PCR inhibitor tests, NFSQ could measure DNA at a concentration of 200 ng/μL of humic acid and 600 μM of hematin. The NFSQ kit showed a DI value trend similar to other qPCR kits. In the reproducibility study, the coefficient of variation of the NFSQ kit was within 10%. The quantitative results of the casework samples obtained using the NFSQ kit were consistent with the STR interpretation results. The NFSQ kit can be useful in the human identification process, as it has detection capabilities similar to those of other comparable quantification kits.

Allele frequencies for 23 autosomal short tandem repeat loci (D3S1358, vWA, D16S539, CSF1PO, TPOX, D8S1179, D21S11, D18S51, TH01, FGA, D5S818, D13S317, D7S820, D2S441, D19S433, D22S1045, D10S1248, D1S1656, D12S391, D2S1338, SE33, Penta D, Penta E), 1 Y-chromosome short tandem repeat locus (DYS391) and Y indel were obtained from 1000 unrelated individuals of the Korean population.

In this study, we compared two Forensic Light Sources (FLSs), LED-based Polilight Flare plus 2 (PF+2) and Laser-based BrightBeam Laser ULTRA (BBL Ultra), in order to detect semen stain from crime scene evidences. The best combinations of FLSs and goggles for detecting semen stain were PF+2/orange and BBL Ultra/yellow, respectively. There were no significant difference in sensitivity for both FLSs. In specificity study, urine, coffee, and juice showed relatively low intensity of fluorescence by both FLSs, however no fluorescence were detected for vaginal fluid, whole blood, and saliva. We found that BBL Ultra is superior to distinguish subtle color and brightness difference between several samples. From the results of background effect tests using semen stain on colored paper, it was shown that PF+2 could not detect the semen stain on the red and pink paper, but BBL Ultra could detect the semen stain. Therefore, we suggested that BBL Ultra was suitable forensic light source for detection of semen stains.

A small and inexpensive thermal cycler (PCR machine), known as the MiniPCR TM Mini8 ThermalCycler (Amplyus, Cambridge, MA, USA), was developed. In this study, the performance of this PCR machinewas compared with the GeneAmp ® PCR system 9700 (Applied Biosystems) using four autosomal short tandemrepeat (STR) kits, a Y-chromosome STR kit, and a mitochondrial DNA HV1/HV2 sequence analysis. Thesensitivity and stochastic effects of the STR multiplex kits and the quality of the DNA sequence analysis weresimilar between the two PCR machines. The MiniPCR TM Mini8 Thermal Cycler could be used for analysesat forensic DNA laboratories and crime scenes. The cost of the PCR is so economical that school laboratoriesand individuals could use the machines.요약: 최근 손안에 들어올 정도로 크기가 작아 범죄현장 등에서 사용이 가능하며, 다른 일반적인 장비들에 비해 가격이 1/10이하로 저렴하여 TM누구나 사용할 수 있는 MiniPCR mini8 Thermal Cycler (Amplyus,Cambridge, MA, USA)가 개발되었다. 본 연구에서는 DNA감식에 일반적으로 사용되고 네 가지 종류의 상염색체 STR 다중증폭 키트들과 한 종류의 Y 염색체 STR 증폭키트, 그리고 미토콘드리아 DNA HV1/HV2의 염기서열 분석법을 사용하여 MiniPCR

A variety of Y-STR analysis kits have been developed and used in the forensic field. Prior to the forensic application of a new kit, laboratory validation and sensitivity tests are essential processes in selecting suitable alternatives and for assuring that standard operating procedures are followed. In this paper, we have performed a sensitivity study of a new commercial kit, the Yfiler ® PLUS PCR Amplification Kit (Yfiler plus kit, released in 2014) by comparing it with the AmpF/STR ® Yfiler TM PCR Amplification Kit (Yfiler kit, released in 2004). The Yfiler plus kit includes the 17 Y-STR loci of the Yfiler kit and has been supplemented with 10 new Y-STR loci. First, we analyzed the sensitivity difference between the two kits using commercial control DNA 2800M and 007. In addition, we compared the detection rate between the two kits from the 16 selected forensic casework samples of less than 0.5 ng concentrations. The results show that the sensitivity and detection rate of the Yfiler plus kit are higher than the corresponding rates of the Yfiler kit. In addition, we were able to obtain more Y-STR profiles with the use of the new kit. Thus, we suggest that Yfiler plus kit is a more effective forensic tool to detect Y-STR profiles from forensic casework samples of low concentrations. 요약 : 법과학 분야에서 다양한 Y-STR 분석 키트가 개발되어 사용되고 있고, 새로운 키트의 법과학적 적용에 앞서 DNA 감정에 적절한 분석 키트들의 선정과 표준작업절차서의 작성을 위해 실험실 내의 내 부적 유효성 검증 및 민감도 시험은 필수적인 과정이다. 본 논문에서는 새로운 상업용 키트인 Yfiler ®

The above article was published with two author names being incorrect. The published paper states "'Hyeun Kyu Yoon and Ki min Seong", whereas it should be "'Hyun Kyu Yoon and Ki Min Seong".

Y chromosome short tandem repeats (Y-STRs) are commonly used to analyze male-specific DNA. Although biallelic patterns due to duplication events have been detected at some loci, Y-STRs generally appear as a single peak except for DYS385 because the Y chromosome is haploid. STR loci in regions of segmental duplication by homologous recombination on the Y chromosome exhibit double allelic peaks, rather than single peaks. In this study, we report a bi- and triallelic pattern observed simultaneously in DYS390, DYS518, and DYS643. A bi- and triallelic pattern has not previously been observed simultaneously for these three loci. We also identified the copy number variation in the region including these loci by the microarray-based analysis. Given the peak balance pattern, the copy number variation, and the close position of these three loci on the Y chromosome, we consider that this phenomenon is caused by a segmental duplication in the euchromatin region. By ruling out mixed samples, a common interpretation of multiple peaks, these results have practical implications for the interpretation of Y-STR results in forensics analyses.

ABO genotyping has been routinely used to identify suspects or unknown remains in crime investigations. Probe-based fluorescence melting curve analysis (FMCA) is a powerful tool for mutation detection and is based on melting temperature shifts due to thermal denaturation. In the present study, we developed a new method for ABO genotyping using peptide nucleic acid (PNA) probe-based FMCA. This method allowed for the simultaneous detection of three single nucleotide polymorphism (SNP) sites in the ABO gene (nucleotide positions 261, 526, and 803) and the determination of 14 ABO genotypes (A/A, A/O01 or A/O02, A/O03, B/B, B/O01 or B/O02, B/O03, O01/O01 or O01/O02 or O02/O02, O01/O03 or O02/O03, O03/O03, A/B, cis-AB01/A, cis-AB01/B, cis-AB01/O01 or cis-AB01/O02, and cis-AB01/cis-AB01). Using this method, we analyzed 80 samples and successfully identified ABO genotypes (A/A [n=5], A/O01 or A/O02 [n=23], B/B [n=3], B/O01 or B/O02 [n=18], A/B [n=9], O01/O01 or O01/O02 or O02/O02 [n=20], cis-AB01/A [n=1], and cis-AB01/O01 or cis-AB01/O02 [n=1]). In addition, all steps in the method, including polymerase chain reaction, PNA probe hybridization, and FMCA, could be performed in one single closed tube in less than 3h. Since no processing or separation steps were required during analysis, this method was more convenient and rapid than traditional methods and reduced the risk of contamination. Thus, this method may be an effective and helpful tool in forensic investigations.

Previously, an age-predictive method based on DNA-methylation patterns in semen was developed, using three CpG sites (cg06304190 in the TTC7B gene, cg12837463, and cg06979108 in the NOX4 gene). Before considering the routine use of a new method in forensics, validation studies such as concordance and sensitivity tests are essential for obtaining expanded and more reliable forensic information. Here, we evaluated a previously described age-predictive method for semen for routine forensic use. Concordance testing showed a high correlation between the predicted and chronological age, with a mean absolute deviation from the chronological age of 4.8 years. Sensitivity testing suggested that age prediction with reliable accuracy and consistency was possible with >5 ng of bisulfite-converted DNA. We also confirmed the applicability of the age-predictive method in forensic casework, using forensic samples. Thus, the proposed method could serve as a very valuable forensics tool for accurate age prediction with semen samples.

DNA methylation has important biological roles, such as gene expression regulation, as well as practical applications in forensics, such as in body fluid identification and age estimation. DNA methylation often occurs in the CpG site, and methylation within the CpG islands affects various cellular functions and is related to tissue-specific identification. Several programs have been developed to identify CpG islands; however, the size, location, and number of predicted CpG islands are not identical due to different search algorithms. In addition, they only provide structural information for predicted CpG islands without experimental information, such as primer design. We developed an analysis pipeline package, CpGPNP, to integrate CpG island prediction and primer design. CpGPNP predicts CpG islands more accurately and sensitively than other programs, and designs primers easily based on the predicted CpG island locations. The primer design function included standard, bisulfite, and methylation-specific PCR to identify the methylation of particular CpG sites. In this study, we performed CpG island prediction on all chromosomes and compared CpG island search performance of CpGPNP with other CpG island prediction programs. In addition, we compared the position of primers designed for a specific region within the predicted CpG island using other bisulfite PCR primer programs. The primers designed by CpGPNP were used to experimentally verify the amplification of the target region of markers for body fluid identification and age estimation. CpGPNP is freely available at http://forensicdna.kr/cpgpnp/.

VNTR D1S80 locus genotyping has been largely replaced in forensics by STR. As the statute of limitations on murder cases was abolished in the Republic of Korea in July 2015, the demand for re-analysis of DNA from unresolved murder cases has increased. The National Forensic Service includes several recorded D1S80 genotypes as crucial clues. Here, we re-established the D1S80 analysis system using capillary electrophoresis and confirmed the reproducibility of the system by comparison with the genotypes of eight DNA samples that had been analyzed using PAGE in 2006. In addition, we created an allelic ladder via new methodology using flanking region sequences. A single DNA sample (K562) and seven primers were used for the new ladder, which contains 12 alleles. Although artificial owing to the use of the flanking region rather than repeat unit reduction, the method is rapid and simple, and could be applicable in any laboratory.

We genotyped and calculated the forensic parameters of 10 non-CODIS loci and 2 CODIS loci of 990 Korean individuals using the InvestigatorⓇ HDplex kit. No significant deviations from Hardy-Weinberg equilibrium (after Bonferroni correction for multiple testing) or genetic linkage disequilibrium were observed. The calculated matching probability and power of discrimination ranged from 0.0080 to 0.2014, and 0.7986 to 0.9920, respectively. We conclude that the markers of the kit are highly informative corroborative tools for forensic DNA analysis.

RUVIS(Reflective Ultraviolet Imaging System) is an effective equipment that detects the location of latent fingerprint at crime scene using short wavelength ultraviolet of 254 nm. In this study, the degree of DNA damage in biological samples was compared depending on the distance and time of processing using four commonly used RUVIS. 50% of DNA was damaged by treating 10 seconds at 10 cm distance in 3 types of RUVIS such as Police RUVIS, SIRCHIE mini light and SIRCHIE RUVIS. In addition, the degree of DNA damage was increased as the distance was closer and the treatment time was longer. It showed that short wavelength UV could cause DNA damage when used close to the samples at crime scene. Therefore, it was suggested to use RUVIS at a distance of at least 1 m. The degree of DNA damage was not significant by Polilight which used long wavelength ultraviolet of 350 nm. As a result, the choice and usage of which UV light and RUVIS were critical for detection of fingerprint and successful DNA typing. 요 약: 범죄 현장에서 눈으로 보이지 않는 지문의 위치를 파악하기 위해 254 nm의 단파자외선과 루비 스(RUVIS;Reflective Ultraviolet Imaging System, 반사자외선이미징시스템) 장비를 사용하는 것이 매우 효과적이다. 최근 유전자 감식 기술의 발전으로 지문과 같은 극미량의 생체시료에서도 성공적으로 DNA 프로필을 확보할 수 있게 되었지만, 지문 탐색에 사용되는 단파자외선에 의해 DNA가 파괴될 수 있다. 본 연구에서는 일반적으로 가장 많이 사용되고 있는 4 종류의 자외선 광원을 대상으로 자외선 조사 시 간과 조사 거리에 따른 DNA 손상 정도를 비교하였다. 단파 자외선을 사용하는 경찰 루비스, SIRCHIE 미니라이트 및 SIRCHIE 루비스의 경우에는 10 cm 거리에서 10초간 조사할 경우 약 50% 정도의 DNA 가 손상되었고, 시료와의 거리가 가까울수록, 처리 시간이 길수록 DNA 손상 정도가 증가하였다. 이 장 비들을 사건 현장에서 사용할 경우에는 유전자 감식 시료의 DNA에 많은 손상을 가져올 수 있기 때문 에 1 m 이상의 거리에서 조사하는 것이 바람직할 것으로 판단되었다. 반면 350 nm의 장파자외선을 사 ★ Corresponding author Phone : +82-(0)33-902-5721 Fax : +82-(0)33-902-5941 E-mail : neobios@korea.kr 148 A-Ram Kim, Woo-Joong Kim, Hee-Yeon Park and Si-Keun Lim Analytical Science & Technology 용하는 폴리라이트 장비는 단파 자외선 장비에 비해 DNA 손상 정도가 크지 않았다. 지문 탐색과 유전 자감식을 모두 고려한다면, 자외선 광원의 종류에 따라 조사 거리와 조사 시간을 결정하는 것이 필요하다.

In this study, we compared two recently released commercial Y-chromosomal short tandem repeat (Y-STR) kits: the PowerPlex Y23 System (PPY23) and Yfiler® Plus PCR amplification kit (YPlus). We performed validation studies, including sensitivity, tolerance to PCR inhibitors, and mixture analysis, and a population genetics study using 306 unrelated South Korean males. PPY23 and YPlus showed similar sensitivity, but PPY23 showed higher tolerance to humic acid than YPlus. Furthermore, the detection rate of unique minor alleles called from male/male mixtures was higher for PPY23 than for YPlus. Comparing the newly added loci, the mean values of gene diversity for PPY23 and YPlus were 0.6715 and 0.8158, respectively. The discrimination capacity in the 306 unrelated South Korean males for PPY23 was 0.9837, and that for YPlus was 0.9935. These results will inform the selection of suitable Y-STR kits based on the purpose of forensic DNA analysis.

The presence of Africans in Britain has been recorded since Roman times, but has left no apparent genetic trace among modern inhabitants. Y chromosomes belonging to the deepest-rooting clade of the Y phylogeny, haplogroup (hg) A, are regarded as African-specific, and no examples have been reported from Britain or elsewhere in Western Europe. We describe the presence of an hgA1 chromosome in an indigenous British male; comparison with African examples suggests a Western African origin. Seven out of 18 men carrying the same rare east-Yorkshire surname as the original male also carry hgA1 chromosomes, and documentary research resolves them into two genealogies with most-recent-common-ancestors living in Yorkshire in the late 18th century. Analysis using 77 Y-short tandem repeats (STRs) is consistent with coalescence a few generations earlier. Our findings represent the first genetic evidence of Africans among 'indigenous' British, and emphasize the complexity of human migration history as well as the pitfalls of assigning geographical origin from Y-chromosomal haplotypes.

The aminoacyl tRNA synthetase complex-interacting multifunctional protein 2 (AIMP2) splice variant designated DX2 is induced by cigarette smoke carcinogens and is often detected in human lung cancer specimens. However, the function of DX2 in lung carcinogenesis is obscure. In this study, we found that DX2 expression was induced by oncogenes in human lung cancer tissues and cells. DX2 prevented oncogene-induced apoptosis and senescence and promoted drug resistance by directly binding to and inhibiting p14/ARF. Through chemical screening, we identified SLCB050, a novel compound that blocks the interaction between DX2 and p14/ARF in vitro and in vivo. SLCB050 reduced the viability of human lung cancer cells, especially small cell lung cancer cells, in a p14/ARF-dependent manner. Moreover, in a mouse model of K-Ras–driven lung tumorigenesis, ectopic expression of DX2 induced small cell and non–small cell lung cancers, both of which could be suppressed by SLCB050 treatment. Taken together, our findings show how DX2 promotes lung cancer progression and how its activity may be thwarted as a strategy to treat patients with lung cancers exhibiting elevated DX2 levels. Cancer Res; 76(16); 4791–804. ©2016 AACR.

We have established 16 small multiplex reactions of two–four loci to amplify 52 recently described single-copy simple Y-STRs and typed these loci in a worldwide panel of 74 diverse men and two women. Two Y-STRs were found to be commonly multicopy in this sample set and were excluded from the study. Of the remaining 50, four (DYS481, DYS570, DYS576 and DYS643) showed higher diversities than the commonly used loci and can potentially provide increased haplotype discrimination in both forensic and anthropological work. Ten loci showed occasional missing alleles, duplicated peaks or intermediate-sized alleles. Electronic supplementary material Supplementary material is available for this article at http://dx.doi.org/10.1007/s00414-006-0124-8 and is accessible for authorized users.

We have identified a Y-chromosomal lineage that is unusually frequent in northeastern China and Mongolia, in which a haplotype cluster defined by 15 Y short tandem repeats was carried by approximately 3.3% of the males sampled from East Asia. The most recent common ancestor of this lineage lived 590 +/- 340 years ago (mean +/- SD), and it was detected in Mongolians and six Chinese minority populations. We suggest that the lineage was spread by Qing Dynasty (1644-1912) nobility, who were a privileged elite sharing patrilineal descent from Giocangga (died 1582), the grandfather of Manchu leader Nurhaci, and whose documented members formed approximately 0.4% of the minority population by the end of the dynasty.

Blood spatter analysis is an important step for crime scene reconstruction. The presence of saliva in blood spatter could indicate expectorated blood which is difficult to distinguish from impact spatter. In this study, four saliva test methods (SALIgAE(®) , Phadebas(®) sheet, RSID(™) -Saliva kit, and starch gel diffusion) were compared to identify the best method for detecting expectorated blood spatter. The RSID(™) -Saliva kit showed the highest sensitivity even when saliva was mixed with blood, and was not inhibited by the presence of blood. The SALIgAE(®) test provided easy and rapid results, but the yellow color of a positive reaction was overwhelmed by the red color of the blood. The starch gel diffusion method and the Phadebas(®) sheet exhibited relatively low sensitivity and the assay took a long time. When using the RSID(™) -Saliva kit for identifying saliva in blood, results should be read within 10 min. Language: en

A small soluble protein, periplasmic tetraheme cytochrome c3, was purified fromDesulphovibrio desulfuricans M6 and its gene, dmc, was cloned and the complete nucleotide sequence determined. The purity index of purified cytochrome c3was 3.3 and the molecular weight was determined as 14.5 kDa by SDS-PAGE. It was found that the 387 bp of dmc gene encoded 21 amino acids of hydrophobic signal peptide and 107 residues of apoprotein. The nucleotide sequence and the predicted amino acid sequence of dmc showed 76% and 83% identities to those of 13 kDa cytochrome c3from D. desulphuricans ATCC 27774, respectively.dmc gene was functionally expressed in aerobically grown Escherichia coli BL-21(DE3) by co-expressing eightccm genes which were reported to be involved in cytochrome c maturation. The molecular weight of overexpressed holocytochrome c3was identical to that of the original protein. Visible spectrum of dithionite-reduced form exhibited typical characteristics of c -type cytochromes. In addition, the redox potential was measured to −340 mV by cyclic voltammetry.