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Article
Internal Validation Guide of the Amp F_STR Identifiler PCR by Using Quantifiler™ Y Human Male DNA Quantification for Use Forensic Laboratories

Author: Mohammed Mahdi AL-Zubaidi
Journal: Al-Nahrain Journal of Science مجلة النهرين للعلوم ISSN: (print)26635453,(online)26635461 Year: 2015 Volume: 18 Issue: 4 Pages: 110-117
Publisher: Al-Nahrain University جامعة النهرين

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Abstract

Fifty samples of buccal swabs were collected from male oral cavity, then genomic DNA was extracted from each sample by two methods, the manual organic phenol chloroform methods and prepfiler forensic extraction kit methods.The concentration of genomic DNA extracted was measured by using Quantifiler® Duo Kit throughout detection the SRY (FAM™-labeled probe), RPPH1(VIC®-labeled probe) and an Internal Positive Control-IPC (NED™-labeled probe).Results showed that the concentration of genomic DNA in buccal swab was ranged between 0.26 and 9.422 ng/ul. these samples work with a serial dilution decimal, as well as PCR .Then introduced to the device 3130xl Genetic Analyzer 16-capillary array system, The results showed that the concentrations is preferable which is located between 25-50 pg, as this did not appear in the concentrations of any problems and the emergence of the peaks clearly Compared with the ALLELIC ladder and positive control, After the election concentrations were chosen Thresholds, Peak amplitude thresholds.

جمعت خمسين عينة من اشخاص اصحاء بواسطة مسحة الفم اجري لها استخلاص بطريقتين الاولى هي التقليدية (فينول كلوروفورم( والطريقة الثانية هي استخدام عدة جاهزة من شركة(Applied Biosystems). بعد استخلاص العينات اجري لها قياس التركيز بواسطة عدة QuantifilerTM human DNA quantitation kit)) التي تتكون من ثلاث بروبات لغرض قياس تركيز DNA. اظهرت نتائج قياس تركيز العينات بانه يتراوح من 0.26-9.422 بيكو غرام. اجريت لها تخافيف عشرية لغرض اختيار التركيز الامثل قبل ادخالها في جهاز الترحيل في الانابيب الشعرية اظهرت النتائج ان افضل تركيز اعطى نتائج خالية من المشاكل التي عادة تصاحب العمل في المجال الجنائي هو من 25-50 بكيوغرام مقارنة مع العينة القياسية. ثم بعدها اختيار خط العتبة الذي ظهرت عندة نتائج جيدة وكذلك اختيار ارتفاع العمود الامثل اذ يمكن اعتمادها في مجال عمل المختبرات الجنائية.


Article
Allele Frequency of 15 Autosomal Short Tandem Repeat (STR) Loci in Al Anbar - Iraqi Population

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Abstract

Abstract:Allele frequencies for 15 STR loci (CSF1PO, D3S1358, D5S818, D7S820, D8S1179, D13S317,D16S539, D18S51, D21S11, FGA, TH01, TPOX, VWA, D2S1338, D19S433) included in the AmpFlSTR Identifiler kit were determined in a sample of 132 unrelated people originating From Al Anbar - Iraqi Population, samples were extracted using a Prep Filer Forensic DNA Extraction Kit. DNA concentration measured using Nano drop Simultaneous amplifications of 15 STR loci and a gender determination marker (multiplexed PCR) were done by using the AmpFlSTR® Identifiler® PCR Amplification Kitaccording to the user’s manual recommendations . The separation and detection of amplified products were conducted with the 3130 xl Genetic Analyzer 16-capillary array system following manufacturer’s protocols,A different number of alleles were observed with frequencies ranging between 0.0038 (FGA- allele 18, 19.2, 23.2, 27 and 28 - , D18S51 - allele 20 and 23, D19S433 - allele 9.2,11 and 12.2, D2S1338 – allele 27, D13S317 – allele 16, D3S1358 – allele 13 and 19 CSF1PO – allele 14, D21S11– allele 30.2 and 32 and D8S1179 - allele 9 and 17) and 0.5795 (TPOX-allele 8). The highest heterozygosity is observed for FGA (85.60 %) where the smallest heterozygosity value is obtained for TPOX (61.36 %). The loci were observed to have high discriminating power, as the power of discrimination of each loci varied from 0.812 (TPOX) to 0.973 (D2S1338). All loci but D21S11 (<0.001), D19S433 (<0.001), D5S818 0.00204 and FGA (<0.00024) met Hardy- Weinberg expectations (P > 0.05).Significant departure from Hardy Weinberg Equilibrium (HWE) expectations were observed in loci D21S11 (0.001), D19S433 (0.001), D5S818 (0.002) and FGA (0.00).The results of the population genetics tests and pairwise comparisons suggest that these allele frequency databases are suitable for the purpose of identification in paternity or forensic investigations.


Article
In Silico and in Vitro Evaluation of Real Time PCR Assay for Detection of Staphylococcus aureus

Authors: Al-Shaimaa Muhammed Saeed Al-Rawi --- Abdul Kareem A. Al-Kazaz --- Majeed Arsheed Sabbah --- Mohammed Mahdi Al-Zubaidi
Journal: Al-Nahrain Journal of Science مجلة النهرين للعلوم ISSN: (print)26635453,(online)26635461 Year: 2017 Volume: 20 Issue: 1 Pages: 126-131
Publisher: Al-Nahrain University جامعة النهرين

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Abstract

Many research groups developed real time PCR assays for identification of Methicillin-resistant Staphylococcus aureus (MRSA). They designed different primers and probes in their assays. Gene mecA is the target identification of MRSA by PCR assay. The aim of this study is using in silico approach to identify the best primers and probe for real time PCR identification mecA gene. Published primers and probes were analyzed in silico to select the best for real time PCR identification of mecA gene. The selected primers and probe successfully used for real time amplification of twenty MRSA tested. This study reveals the importance of in silico approach for designing diagnostic assays shorten the cost and the time. [DOI: 10.22401/JNUS.20.1.18]

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