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A comparative study of ARMS-PCR and RFLP-PCR as methods for rapid SNP identification

Duţă-Cornescu G, Simon-Gruiţă A, Constantin N, Stanciu F, Dobre M, Banică D, Tuduce R, Cristea P, Stoian V. Romanian Biotechnological Letters, 2009;14(6):4845-4850.

Abstract:

Identification of single nucleotide polymorphisms (SNPs) is now possible by many techniques, but choosing one of these methods for a particular case represents quite a challenge, because the researcher must take into consideration many factors.
In this article the authors are trying to present a comparative study of two methods, used currently in our laboratory, for identification of SNPs polymorphisms: ARMS – PCR (amplification refractory mutation system) and RFLP – PCR (restriction fragment length polymorphism). The two SNPs on which we focused belong to human VDR gene (vitamin D receptor gene) and are ApaI (a G→T base change in intron 8) and TaqI (a silent T→C base change in codon 352), named after the restriction enzymes which recognize these variations.
Since the results obtained by both methods were confirmed by direct sequencing, we concluded that ARMS-PCR method is the most adequate for detecting the alternative genotypes determined by single base mutations. The simplicity of this method makes it suitable for the analysis of large number of samples, situation which is usually met in case-control and population genetic studies because this test is easy to use, cost – effective and have an accuracy of 99,9%

Keywords:ARMS-PCR, RFLP-PCR, sequencing, VDR gene, Apa I and Taq I polymorphisms

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Allele frequencies of 15 STR loci in Moldavia region (NE Romania)

Stanciu F, Popescu OR, Stoian IM. Forensic Sci Int Genet. 2009 Dec;4(1):e39-40. doi: 10.1016/j.fsigen.2009.02.008. Epub 2009 Apr 8. PMID: 19948322.

Abstract:

Allele frequencies for 15 STR loci included in AmpFlSTR Identifiler kit (D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, vWA, TPOX, D18S51, D5S818, and FGA) were determined in a sample of 1321 unrelated iniduals living in the region of Moldavia (NE Romania). No deviations from Hardy–Weinberg equilibrium were observed (only after applying the Bonferroni correction in the cases of D2S1338). Genetic parameters of forensic interest were calculated and comparison with geographically nearby populations was performed.

Keywords:DNA typing, Short tandem repeats (STRs), AmpFlSTR Identifiler, Population data, Moldavia

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Automated DNA isolation for databasing purposes

Stanciu F, Stoian IM, Popescu OR. Rom J Leg Med 2009;17(1):51–58

Abstract:

Nowadays, automation is crucial in databasing laboratories especially for countries which are relative new in the field and must quickly reach the minimum standards of consecrated databasing laboratories from Europe and USA. In our efforts to supply genetic profiles for Romanian National DNA Database, we evaluated the Freedom Evo 150 liquid handling platform with AGOWA sep9600 magnetic separator in different running conditions and workflows, for reaching the most efficient ones. We compared the results with those obtained from manual DNA isolation with Chelex resin and paramagnetic particles. For this we used a total of 6246 biological reference samples (5220 – automated processed and 1026 – manually processed), amplified using AmpFlstr Identifiler Kit and analyzed by capillary electrophoresis using ABI Prism 3100 Genetic Analyzer.

Keywords: automation, databasing laboratory, liquid handling platform, DNA magnetic separator.

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STR data for the AmpFlSTR Identifiler from Dobruja region (SE Romania)

Stanciu F, Popescu OR, Stoian IM. Forensic Sci Int Genet. 2009 Mar;3(2):146-7. doi: 10.1016/j.fsigen.2008.09.009. Epub 2008 Oct 26. PMID: 19215887.

Abstract: 

Allele frequencies for 15 STR loci included in AmpFlSTR Identifiler kit (D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, vWA, TPOX, D18S51, D5S818, and FGA) were determined in a sample of 569 unrelated iniduals living in the region of Dobruja (SE Romania). No deviations from Hardy–Weinberg equilibrium were observed. Genetic parameters of forensic interest were calculated and comparison with geographically nearby populations was performed.

Keywords: DNA typing, Short Tandem Repeats (STRs), AmpFlSTR Identifiler, Population

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Population Data for 15 Short Tandem Repeat Loci from Wallachia Region, South Romania

Stanciu F, Stoian IM, Popescu OR. Croat Med J. 2009 Jun;50(3):321-5. doi: 10.3325/cmj.2009.50.321. PMID: 19480027; PMCID: PMC2702745.

Aim: To determine allele frequencies’ distribution for 15 short tandem repeat (STR) loci in a population sample of 1910 unrelated iniduals from the region of Wallachia, South Romania.

Methods: DNA was isolated using Chelex 100 method and an adapted version of AGOWA mag DNA Isolation Kit Sputum. Polymerase chain reaction amplification was done using AmpFlSTR Identifiler kit (D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, vWA, TPOX, D18S51, D5S818, and FGA). For DNA typing, ABI PRISM 3100 Genetic Analyzer was used. Genetic parameters of forensic interest were calculated and comparisons with geographically close populations were performed.

Results: With the exception of vWA locus (P=0.001), no other significant deviations from Hardy-Weinberg expectations were found. Single locus comparisons with data on geographically close populations showed significant differences between the population of Wallachia and the population of Bucharest area, Greece, Turkey, Italy, Hungary, Belarus, and Poland, but no differences were found from the population from Croatia and Serbia.

Conclusion: According to 15 analyzed STR loci, the population of Wallachia region was found to be genetically more similar to Slavic populations of Croatia and Serbia than to other surrounding populations.

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