scholarly journals Assessment of QIAGEN™ Investigator® 24plex GO! kit workflow for autosomal STR profiling of forensic reference samples

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Hashom Mohd Hakim ◽  
Hussein Omar Khan ◽  
Siti Afifah Ismail ◽  
Nurul Hazirah Mat Lazim ◽  
Japareng Lalung ◽  
...  
Keyword(s):  
Pcr Amplification ◽  
Dna Profiling ◽  
Buccal Swab ◽  
Forensic Dna ◽  
Str Typing ◽  
Str Analysis ◽  
Dna Database ◽  
Autosomal Str ◽  
Dna Template ◽  
Reference Samples

Abstract Background DNA profiling has proven to be a valuable technique for identification of individuals in crime. Currently, the technique targets several short tandem repeat (STR) regions in human genome. However, increasing number of samples submitted for STR analysis may lead to delays due to the limited number of experienced analysts who might be available at any given moment and the time taken to complete lengthy DNA profiling procedures. This study was conducted to test the specificity, repeatability, reproducibility and robustness of Investigator® 24plex GO! kit for genotyping of reference samples submitted to the Royal Malaysian Police Forensic DNA Laboratory for DNA database. Material and methods In this study, Investigator® 24plex GO! kit was used to directly amplify STR loci from buccal swab cell of reference samples that had previously been STR typed using GlobalFiler™ Express kit. Capillary electrophoresis was carried out on a 3500xL Genetic Analyser using POP-4® Polymer. Amplified products were assigned to particular STR alleles using the GeneMapper ID-X version 1.4 software. Results Our study shows that STR profiles generated using Investigator® 24plex GO! gave concordance results with those previously obtained using the GlobalFiler™ Express kit. In addition, quality sensors included in the kit are of particular importance for determining the effectiveness of the PCR reaction and help to indicate the nature and quantity of DNA template for PCR amplification. Conclusion The Investigator® 24plex GO! kit is reliable for STR typing of reference samples.

scholarly journals Assessment of autosomal and male DNA extracted from casework samples using Casework Direct Kit, Custom and Maxwell 16 System DNA IQ Casework Pro Kit for autosomal-STR and Y-STR profiling

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Hashom Mohd Hakim ◽  
Hussein Omar Khan ◽  
Siti Afifah Ismail ◽  
Shahrizad Ayob ◽  
Japareng Lalung ◽  
...  
Keyword(s):  
Tandem Repeats ◽  
Pcr Amplification ◽  
Dna Profiling ◽  
Nucleic Acid Content ◽  
Str Profiling ◽  
Autosomal Str ◽  
Y Chromosomes ◽  
Y Chromosome Str ◽  
Equal Capacity ◽  
Dna Template

Abstract Short repetitive regions in autosomal and Y chromosomes known as short tandem repeats (STRs) are currently used for DNA profiling in crime investigations. However, DNA profiling requires a sufficient quality and quantity of DNA template, which is often not obtained from trace evidence or degraded biological samples collected at the scene of a crime. Here, we assessed autosomal and male DNA components extracted from crime scene and mock casework samples using the Casework Direct Kit, Custom and compared the results against those obtained by extraction of matching samples using well-established Maxwell 16 System DNA IQ Casework Pro Kit. The quantity and quality of extracted DNA obtained using both Casework Direct Kit, Custom and Maxwell 16 System DNA IQ Casework Pro Kit were analyzed using PowerQuant Systems followed by autosomal and Y-chromosome STR profiling using GlobalFiler Express PCR Amplification Kit and PowerPlex Y23 System, respectively. Our results showed that the Casework Direct Kit and Maxwell 16 DNA IQ Casework Pro Kit have more or less equal capacity to extract inhibitor free DNA, but that the latter produces slightly better quality and more DNA template and subsequently higher numbers of STR allele calls for autosomal and Y-STR analyses. Nonetheless, the Casework Direct Kit, Custom is the quicker and cheaper option for extraction of good, clean DNA from high content material and might best be used for extraction of reference samples. Such reference DNA samples typically come from buccal swabs or freshly drawn blood. So, in general, they can confidently be expected to have a high nucleic acid content and to be inhibitor-free.

scholarly journals Optimisation of rapid STR analysis using a standard DNA forensic pipeline

2019 ◽  
Author(s):  
Katharine Gammon ◽  
Carl Mayers
Keyword(s):  
Capillary Electrophoresis ◽  
Pcr Amplification ◽  
Buccal Cell ◽  
Forensic Dna ◽  
Direct Pcr ◽  
Str Analysis ◽  
Dna Profile ◽  
Fta Cards ◽  
Dna Profiles ◽  
Reference Samples

Previous studies in published literature have reported on various alterations to STR mastermixes, protocols and instrumentation in order to reduce the time taken to generate forensic DNA profiles from reference and casework type samples. In this study, we demonstrate how altering default PCR amplification and capillary electrophoresis protocols in our existing DNA profiling pipeline can reduce the overall time taken to generate a DNA profile from buccal cell reference samples. GlobalFiler Express STR mastermix was used with direct PCR from FTA cards, run on altered PCR protocols and CE settings, and results compared to the standard evaluated settings used in our laboratories. This study demonstrated that full DNA profiles could be recovered in less than 80 minutes in comparison to our standard time of 97 – 102 minutes whilst utilising existing reagent kits and instrumentation, with only minor modifications to protocols.

scholarly journals Validation Study of 6 Dye-Based SF25TM PCR Amplification Kit

2021 ◽  
Author(s):  
Pankaj Shrivastava ◽  
R. K. Kumawat ◽  
Pushpesh Kushwaha ◽  
Akshay Kumar ◽  
Manisha Rana ◽  
...  
Keyword(s):  
Dna Analysis ◽  
Pcr Amplification ◽  
Turnaround Time ◽  
Fluorescent Dye ◽  
Forensic Dna ◽  
Allelic Ladder ◽  
Forensic Dna Analysis ◽  
Inhibitor Tolerance ◽  
Autosomal Str ◽  
Dna Template

Abstract Ever since the inception of STR-based forensic DNA analysis for human Identification and DNA profiling, the approach developed continuously. Most of these developments are pertaining to achieve the best polymorphic loci set for analysis, high inhibitor tolerance capability, increase in sensitivity, reduction in turnaround time, and cost-effectiveness. On the similar line, the SF25TM PCR amplification kit, a 6 fluorescent dye-chemistry based autosomal STR kit was developed, which analyses 25 STR loci viz., FAM fluorescent dye-labelled D3S1358, D13S317, D7S820, D16S539, D1S1656, and Penta E; HEX fluorescent dye-labelled TPOX, TH01, D2S1338, CSF1PO, and Penta D; SUM fluorescent dye-labeled D19S433, vWA, D21S11, D18S51, and D6S1043; LYN fluorescent dye-labelled Amelogenin, D8S1179, D5S818, D12S391, and FGA; PUR fluorescent dye-labelled D22S1045, Y indel; D2S441 and D10S1248, The developmental validation of the SF25 PCR Amplification reagents were carried out in this study to analyze its specificity, sensitivity, and concordance. This multiplex kit showed higher sensitivity with 125pg of DNA template generating a complete profile (with 100% allele call). Out of 83 samples, 10 samples showed aberrant triallelic pattern/aberrant heterozygous pattern at D16S539 locus. The possible discrepant homozygous condition at locus D1S1656 might be due to the invasion of alleles at D1S1656 locus present adjacent to D16S539 locus. Such invasion resulted due to the shortening of the allelic bin at the D1S1656 locus. The allelic ladder of the SF25TM PCR amplification kit reveals the allelic range of D1S1656 from 11 to 19.3 which can be interpreted according to the casework as deemed by the end-users for their samples.

scholarly journals ISOLASI FRAGMEN GEN PENYANDI PUTRESIN N-METILTRANSFERASE DAN QUINOLINAT FOSFORIBOSILTRANSFERASE ASAL TEMBAKAU LOKAL TEMANGGUNG (Nicotiana tabacum)

2020 ◽  
Vol 17 (3) ◽  
pp. 109 ◽  
Author(s):  
SESANTI BASUKI ◽  
NURHAJATI AA MATTJIK ◽  
SUWARSO SUWARSO ◽  
DESTA WIRNAS ◽  
SUDARSONO SUDARSONO
Keyword(s):  
Dna Sequences ◽  
Pcr Amplification ◽  
Data Bank ◽  
Degenerate Primer ◽  
Gene Encoding ◽  
Methyl Transferase ◽  
Dna Database ◽  
Genes Encoding ◽  
Tobacco Cultivar ◽  
Dna Template

<p>ABSTRAK</p><p>Upaya untuk menurunkan kandungan nikotin merupakan salah satuprioritas utama penelitian tembakau. Nikotin adalah senyawa alkaloidutama berpotensi dikonversi menjadi senyawa nor-nikotin yang bersifatkarsinogen. Gen PMT sebagai penyandi enzim putresin n-metiltransferase(PMT) dan gen QPT - penyandi enzim quinolinat fosforibosiltransferase(QPT) merupakan dua gen kunci yang berperan penting pada proses bio-sintesis nikotin. Penelitian ini bertujuan untuk mengisolasi potongan genPMT dan QPT asal tembakau lokal Indonesia, mengkarakterisasi danmenganalisis runutan DNA-nya. Tahapan penelitian dimulai dengan me-rancang primer degenerate berdasarkan informasi yang ada di pangkalandata Bank Gen NCBI (National Centre for Biotechnology Information),mengamplifikasi PCR menggunakan templat DNA genomik tembakaulokal cv. Sindoro1, mengklon potongan DNA hasil PCR dan menentukanrunutan DNA-nya. Hasil penelitian menunjukkan dari dua belas pasangprimer degenerate yang dirancang, hanya dua pasang primer yang meng-hasilkan potongan DNA hasil amplifikasi PCR, yaitu pasangan primerPMt-7 (F &amp; R) untuk gen PMT dan primer QPt-3 (F &amp; R) untuk gen QPT.Setelah dilakukan penentuan runutan DNA-nya, amplikon yang didapatdari hasil PCR dengan pasangan primer PMt-7 sebesar 1418 bp, sedangkanuntuk primer QPt-3 sebesar 205 bp. Runutan DNA gen PMT dan gen QPTasal tembakau lokal cv. Sindoro1 mempunyai tingkat kesamaan yang ting-gi dengan gen PMT dan gen QPT asal tembakau lainnya yang ada dipangkalan data Bank Gen NCBI.</p><p>Kata kunci : Gen PMT, gen QPT, lintasan biosintesis nikotin, perunutanDNA, amplifikasi PCR, primer degenerate</p><p>ABSTRACT</p><p>Isolation of Genes encoding Putrescine N-Methyl-transferase and Quinolinat Phosphoribosyl transferasederived from Temanggung Tobacco Cultivar (Nicotianatabacum)</p><p>Reduction of nicotine content is one of the major objective intobacco research. Nicotine is the main alcaloid compound that potentiallycould be converted into a carcinogenic compound (nor-nicotine). The PMTgene encoding putrescine N-methyl transferase (PMT) and the QPT gene -encoding quinolinate phosphoribosyl transferase (QPT) are the two keyenzymes involved in nicotine biosynthesis. The objectives of this researchwere to isolate PMT and QPT gene fragments originated from Indonesianlocal tobacco, to characterize, and to analyze their DNA sequences. Theresearch activities included: degenerate primer design based oninformation available in the GenBank DNA Database NCBI (NationalCentre for Biotechnology Information), PCR amplification usingdegenerate primer and genomic DNA template of a local tobacco cv.Sindoro1, clone the PCR amplified products, and determine their DNAnucleotide sequences. Results of the experiment indicated that from 12degenerate primer pairs synthesized, only two were able to yield positivePCR amplified products. These primer pairs were PMt-7 (F &amp; R primers)for PMT and QPt-3 (F &amp; R primers) for QPT. After DNA sequencing, theamplified DNA product amplified using PMt-7 degenerate primer pairswere 1418 bp, while that using QPt-3 primer pairs were only 205 bp.Nucleotide sequences of PMT or QPT gene fragments originated fromlocal tobacco cv. Sindoro1 showed a high nucleotide sequences identity ascompared to that of the respective genes from other tobacco species thatwere available in the GenBank DNA Database NCBI.</p><p>Key words: PMT gene, QPT gene, nicotine biosynthetic pathways, DNAsequencing, PCR amplification, degenerate primer</p>

scholarly journals Efficient DNA Sampling in Burglary Investigations

Genes ◽  
2021 ◽  
Vol 13 (1) ◽  
pp. 26
Author(s):  
Colin Charles Tièche ◽  
Markus Dubach ◽  
Martin Zieger
Keyword(s):  
Crime Scene ◽  
Forensic Dna ◽  
Dna Profile ◽  
Dna Database ◽  
Dna Sampling ◽  
High Incidence ◽  
Crime Scene Investigators ◽  
Dna Collection ◽  
Private Homes ◽  
Reference Samples

In terms of crime scene investigations by means of forensic DNA-analyses, burglaries are the number one mass crime in Switzerland. Around one third of the DNA trace profiles registered in the Swiss DNA database are related to burglaries. However, during the collection of potential DNA traces within someone’s residence after a burglary, it is not known whether the sampled DNA originated from the perpetrator or from an inhabitant of said home. Because of the high incidence of burglaries, crime scene investigators usually do not collect reference samples from all the residents for economical and administrative reasons. Therefore, the presumably high probability that a DNA profile belonging to a person authorized to be at the crime scene ends up being sent to a DNA database for comparison, has to be taken into account. To our knowledge, no investigation has been made to evaluate the percentage of these non-perpetrator profiles straying into DNA databases. To shed light on this question, we collected reference samples from residents who had been victims of recent burglaries in their private homes. By comparing the profiles established from these reference samples with the profiles generated from trace DNA, we can show that the majority of the DNA samples collected in burglary investigations belong to the residents. Despite the limited number of cases included in the study, presumably due to a crime decline caused by the pandemic, we further show that trace DNA collection in the vicinity of the break and entry area, in particular window and door glasses, is most promising for sampling perpetrator instead of inhabitant DNA.

scholarly journals Interpretation of DNA data within the context of UK forensic science — evaluation

2021 ◽  
Author(s):  
Roberto Puch-Solis ◽  
Susan Pope
Keyword(s):  
State Of The Art ◽  
Crime Scene ◽  
Database Search ◽  
Dna Profiling ◽  
Forensic Dna ◽  
Criminal Trials ◽  
Kinship Analysis ◽  
Dna Database ◽  
Forensic Dna Profiling ◽  
The Uk

Forensic DNA provides a striking contribution to the provision of justice worldwide. It has proven to be crucial in the investigative phase of an unsolved crime where a suspect needs to be identified, e.g. from a DNA database search both nationally and internationally. It is also a powerful tool in the assignment of evidential weight to the comparison of a profile of a person of interest and a crime scene profile. The focus of this document is the evaluation of autosomal profiles for criminal trials in the UK. A separate review covers investigation and evaluation of Y-STR profiles, investigation using autosomal profiles, kinship analysis, body identification and Forensic Genetic Genealogy investigations. In less than 40 years, forensic DNA profiling has developed from a specialist technique to everyday use. Borrowing on advances in genome typing technology, forensic DNA profiling has experienced a substantial increase in its sensitivity and informativeness. Alongside this development, novel interpretation methodologies have also been introduced. This document describes the state of the art and future advances in the interpretation of forensic DNA data.

scholarly journals Sperm Cell Capture Based on ABH Antigen Differences to Separate Two Men in Mixed Seminal Stains

2021 ◽  
Vol 2021 ◽  
pp. 1-5
Author(s):  
Mao-ling Sun ◽  
Ji-long Zheng ◽  
Bao-jie Wang ◽  
Jun Yao
Keyword(s):  
Sperm Cell ◽  
Tandem Repeats ◽  
Pcr Amplification ◽  
Personal Identification ◽  
Blood Type ◽  
Cell Capture ◽  
Str Typing ◽  
Autosomal Str ◽  
Unrelated Donors ◽  
Seminal Stains

Personal identification of two individuals in mixed semen samples in forensic DNA testing in general usually involves analysis using autosomal and Y chromosome short tandem repeats (STRs). Results may exclude unrelated donors but cannot identify individuals. In this study, sperm cell capture based on ABH antigen differences was used to obtain the cells with the single ABO blood type. Immunohistochemical staining using labeled anti-A, anti-B, and anti-H antibodies and the laser microdissection system can be used to enrich sperm with different ABO types in mixed seminal stains from two individuals. Then, PCR amplification and capillary electrophoresis were performed to genotype the STR loci. To some extent, after sperm cell capture based on ABH antigen differences, autosomal STR typing using enriched single blood group cells can be utilized to partially identify different individuals in a mixed seminal stain sample from two individuals.

scholarly journals Awareness level on the role of forensic DNA database in criminal investigation in Nigeria: A case study of Benin city

2020 ◽  
Vol 4 (1) ◽  
pp. 007-014
Author(s):  
Udogadi Nwawuba Stanley ◽  
Blessing Nkiruka Akpata Chinyere
Keyword(s):  
Dna Profiling ◽  
Forensic Dna ◽  
Chi Square ◽  
Statistical Tool ◽  
Benin City ◽  
Dna Database ◽  
Forensic Dna Profiling ◽  
Awareness Level

Pieces of evidence have continued to emerge, demonstrating the extensive efficiency and effectiveness of the DNA database in assisting criminal investigations around the world. Therefore, the present study aimed to determine the awareness level on the prominent role of Forensic DNA Database on Crime Investigation in Nigeria: a case study of Benin City. In conducting this research, a total of 458 questionnaires were distributed around Benin City between the periods of 12th January 2020 to 21st March 2020, with a particular focus on security agents and students. The questionnaire comprised of three main categories: Socio-demographic characteristics, Information about the National Forensic DNA Database, and Information about DNA evidence, and Nigeria Criminal Justice system. For the analysis of data collected; the statistical tool used was also Statistical Package for Social Sciences, version 22 for windows. Responses were compared using chi-square and presented as counts and percentages. In determining the level of awareness, the following responses were obtained. Of the total population: 53.28% had no idea about forensics, 19.21% were uncertain and 27.54% knew about forensics. The same trend was observed with Forensic DNA profiling, 42.14% did not know, 22.27% were uncertain and 35.59% demonstrated good knowledge of Forensic DNA profiling. On the knowledge about the National Forensic DNA Database, 48.47% had no knowledge, 22.27% were uncertain and 29.26% were knowledgeable about it. The result of the present study revealed that the awareness level of the forensic DNA Database was found to be inadequate.

Sign in / Sign up

Export Citation Format

Share Document