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Öğe Comparison and Development of a Rapid Extraction Method of DNA From Ancient Human Skeletal Remains of Turkey(2009) Vural, H. C.; Tırpan, A. A.The use of genetic technology in forensic science is applied primarily to distinguish between individials who may be the source of biological material associated with archeological remains. DNA sequences from ancient fossils have great potential for studies of phylogeny, biogeography and molecular evolution. DNA from fossils also facilitates the rigorous testing and calibration of mutation rates among related taxa, sex test and molecular divergence time. In this study, a rapid and quantitative aDNA extaction methods from human skeletal remains was developed for application of forensic science and archeometry. For that reason, DNA was extracted from ancient human bones from Mugla in Turkey. Extraction of DNA was carried out using the laboratuary handling and cleaning protocol. After cleaning of bone, small piece of ancient bones were ground to powder with a mixer mill. Aliquots of the powder were subjected to a calfication method and extracted with 0.5 M EDTA (pH 8.3) for 48 hours at 56 C. After addition of proteinase K, solution of bone was incubated at 37 C. Genomic DNA from supernatant was extracted automatically by using EZ1 Automatic Nucleic Acid Isolation System (Qiagen, Germany) with investigator kit (Qiagen, Ilden, Germany) and different DNA extraction methods which are modified by researcher from ancient bones. EZ1 Nucleic acid isolation method; This tehnique is quite useful for high yield and quality of aDNA isolation from human skeletal remains. In this methods, no further purification was needed for molecular analysis. Amount and purity of extracted DNA from ancient bones were measured by Spectrophotometer. In addition to spectrophotometric measurement, extracted DNA was applied to 1% agarose gel, stained and imaged under ultraviolet (UV) irradiation. As a result, 50 ng pure DNA was extracted from ancient bones, approximately 1.8. This protocol proved to be advantageous because of its simplicity, quickness and affordable reagents, besides the high molecular weight DNA and purity achieved in a variety of fosil bone tissues from the total set obtained from Mugla in Turkey.Öğe Comparison of winter and summer canola (Brassica napus) genotypes in Turkey(FUNPEC-EDITORA, 2013) Maltas, E.; Vural, H. C.We examined genetic relationships between canola (Brassica napus) genotypes cultivated in winter and spring in Turkey. Genomic DNA was isolated from the seeds by two modified CTAB protocols: EZ1 nucleic acid isolation method and a commercial kit (Dneasy Plant Mini Kit, Qiagen). Diversity and genetic relationships in the genotypes were analyzed with RAPD markers; 156 reliable bands were found for both genotypes, of which 24% were polymorphic. Fifteen primers gave at least one consistent polymorphic band. The dendogram developed by pooling data of RAPD analysis of summer and winter genotypes had similar patterns. This technique allowed us to examine the relationship between canola genotypes.Öğe Genetic Identification of Soybean [Glycine Max (L.) Merr.] Growing in Turkey for Molecular Breeding Using Molecular Markers(TAYLOR & FRANCIS LTD, 2010) Vural, H. C.The Soybean (Glycine max) is a species of legume which is a plant in the family Fabaceae. It is an annual plant that has been used in China and in other countries for 5 000 years as a food and a component of drugs. Soy contains significant amounts of all the essential amino acids for humans, and therefore is a good source of protein. Soybeans are the primal:), ingredient in many processed foods, including daily product substitutes. The feasibility of detecting yields quality in soybean genotypes by a polymerase chain reaction (PCR) method is determined. PCR is a sensitive method for analyzing DNA and it is considered the most important method for detection of soybeans in processed or raw foods. Namely, PCR is a commonly applied nucleic amplification method which is specific and sensitive enough to detect even tiny amounts of organism-specific DNA sequences. This study focuses on the PCR detection in food, describes rapid and reliable DNA extraction methods which can be applied to a variety of food samples and details of PCR amplification protocols for sensitive and specific detection of soybean genotypes growing in Turkey. All soybean samples were evidenced by all PCR primers as soybean products.Öğe Molecular Analysis of Chickpea Species Through Molecular Markers(DIAGNOSIS PRESS LTD, 2010) Vural, H. C.; Akçin, A.Chickpea is an important food legume crop with high nutritional value. Lack of suitable DNA isolation protocol is a limiting factor for any molecular studies on this crop. Today, however; ills possible to determine plants that possess desired qualities in earlier generations using molecular methods (DNA marker techniques) and combine gene areas bearing the desired qualities in commercially used varieties. Therefore, in later studies, 5 ISSR primers, which were pre-determined in chickpea genome mapping and characteristic of resistance, were tested on 10 species (C. arietinum, C. reticulatum, C. pinnatifidum, C. anatolicum, C. echinospermum, C. bijugum, C. judaicum, C. yamashitae, C. chorassanicum, C. soongaricum) in order to study quantitative traits of the crop, a polymorphism was determined among the species and it was proven that various PCR (Polymerase Chain Reaction) parametres were important in achieving amplification with molecular markers. The present study preferred the ISSR marker because, in studies based on PCR, it is economical and practical to obtain definitive results at the shortest time and such that are being used in routine studies. Besides, the study also found that by virtue of its high incidence of polymorphism and supply of amplification.Öğe RT-qPCR assay on the vitamin D receptor gene in type 2 diabetes and hypertension patients in Turkey(FUNPEC-EDITORA, 2012) Vural, H. C.; Maltas, E.RT-qPCR was used to analyze the vitamin D receptor (VDR) gene TaqI polymorphism in 100 Turkish patients with type 2 diabetes mellitus (T2DM) and hypertension compared with 100 healthy subjects, to determine whether VDR could be considered as one of the susceptibility genes for T2DM and hypertension. Genotyping was done with PCR, followed by melting curve analysis with specific fluorescent hybridization probes. The results showed that distributions for TT, Tt and tt genotypes were 51, 46 and 3% in the patient group, and 35, 49 and 16% in the control group, respectively. The frequency of the T allele in patients was also significantly higher than that in controls. Based on the results, the relationship between the VDR gene TaqI polymorphism and T2DM patients in the Turkish population was compared. In terms of the genotype distributions and allele frequencies of the VDR gene TaqI polymorphism, there was no statistically significant difference (P > 0.05) between the T2DM and hypertension patients and controls. Application of RT-qPCR method enabled us to assess the prevalence of the VDR gene TaqI polymorphism and its association with type 2 diabetes and hypertension.Öğe Study on Polymorphism and Activities of GSTM1 and CYP1A1 Genes in Connection with Various Cancer Types in Turkey Population(TAYLOR & FRANCIS LTD, 2010) Vural, H. C.; Turaclar, N.; Elagöz, S.The metabolism of drugs and chemical carcinogens involves a variety of isoenzymes such as members of the Cytochrome P450 (CYP) and Glutathione S-transferase (GSTs) families, with polymorphisms described in these genes appearing to be responsible for differences in individual susceptibility to cancer Furthermore, the cytochrome P450 family (CYPs) and the glutathione S-transferase (GSTs) enzymes also play an important role in the metabolism of environmental carcinogens and of estrogen and can affect various cancer types. In this study we examine the role of the genes CYP1A1 and GSTM1, in different cancer types in Turkey population. The aim of our work was to evaluate the association between CYP1A1 and GSTM1 genetic polymorphisms and activities of genes indicative for susceptibility to various cancer types in Turkey population. The polymorphic inheritance of human drug-metabolizing enzymes, such as those encoded by the GST and CYP systems, has been implicated in all the cancer risk and prognostic. The study population consisted of 80 different incident cancer cases and 100 healthy controls. Genotyping analyses were performed by PCR based methods. In conclusion, in our population GSTM1 was associated with breast, gastric, endometrium, testis, parotis, lymph node, pancreas, tyroid, and lung cancer risk in Turk population. The analysis of patients by histological types of different cancer showed no association between histopathologic types of cancer and CYP1A1 gene polymorphism (p=0.6). Genetic researches using specific biomarkers are expected to be helpful in evaluation the risks for every cancer type.
