Abstracts / Journal of Biotechnology 208 (2015) S5–S120
with 5 and 3 primers on each side of the mutation. The 237bp-PCR product is digested with Taq I restriction endonuclease for two hours at 37 ◦ C. The fragments obtained after digestion are separated by using electrophoresis in 10% polyacrylamide gel. For the detection of Asn291Ser mutation, exon 6 is amplified by using the PCR technique with a 5 -PCR primer complementary to a sequence located in intron 5 near the 5 boundary of exon 6 and a 3 -“mismatch” PCR primer complementary to a sequence located in exon 6 near the Asn291Ser mutation.The 238-bp-PCR product is then digested with Rsal restriction endonuclease, for two hours at 37 ◦ C. The digested fragments are separated by electrophoresis on 2% agarose gel. The research into the presence of these two mutations on LPL gene will generate essential elements in monitoring and managing patients with metabolic syndrome. http://dx.doi.org/10.1016/j.jbiotec.2015.06.262 The RFLP profiles at BRAF V600E mutations in thyroid FNAB nodules Semra Ozdemir 1,∗ , Mehmet Asık 3 , Fatma Silan 2 , Ozturk Ozdemir 2 , Yusuf Ziya Tan 1 , Elif Ari 2 , Mustafa Eroglu 3 , Kubilay Ukinc 3 1
Canakkale Onsekiz Mart University, Faculty of Medicine, Department of Nuclear Medicine, 17100 Canakkale, Turkey 2 Canakkale Onsekiz Mart University, Faculty of Medicine, Department of Medical Genetics, 17100 Canakkale, Turkey 3 Canakkale Onsekiz Mart University, Faculty of Medicine, Department of Endocrinology and Metabolic Disease, 17100 Canakkale, Turkey
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our clinic with mutism without hearing problems and hyperactivity. 30 years old mother had a high risk of neural tube defect in the triple screen test. Physical examination revealed hidden penis and underdevelopment of the scrotum. Radiology observed testes in inguinal canals. Psychometric tests showed a mild developmental delay and a serious deficiency in speaking ability. Results of cytogenetic analysis showed 46, XX subsequently the next step was the determination of SRY gene by FISH, and AZF gene by multiplex Flourescent PCR method. SRY gene signal, which is normally located on Yp11.3, was observed on the p terminal site (Xp22.3) of one of the X chromosomes [46,XX, + mar ish.add (X pter) (wcpY + SRY)]. AZF gene could not be determined by multiplex Flourescent PCR method, but fluorescent signals for SRY and ZFY genes, which are control for this analysis, were also positive. Despite father’s blood test is negative, gonadal mosaicism is an option, too. Also during gametogenesis while crossing over between pseudoautosomal regions there is a chance of translocation of SRY and loss of genetic materials which is resulted in disabilities of this patient. http://dx.doi.org/10.1016/j.jbiotec.2015.06.264 The microdeletion/microduplication profiles in spontaneously aborted fetal materials: Double blind results of QF-PCR and MLPA techniques Fatma Silan, Elif Ari, Ahmet Uludag, Onur Yildiz, Betul Isin, Baris Paksoy, Mine Urfali, Ozturk Ozdemir ∗ Department of Medical Genetic, Faculty of Medicine, Canakkale Onsekiz Mart University, 17100 Canakkale, Turkey
E-mail address:
[email protected] (S. Ozdemir).
E-mail address:
[email protected] (O. Ozdemir).
It was aimed to investigate the prevalence of the BRAF V600E mutation in FNAB of thyroid nodules. The study consists of a total of 51 patients who had cold thyroid nodule in Tc-99 m thyroid scintigraphy. FNAB were obtained from hypoactive nodules and total genomic DNA were isolated. DNA samples were used for PCR amplifications and all amplicons were genotyped by RFLP technique for target BRAF V600E polymorphism. Solute tumors were genotyping by RFLP tecnique by usening TSP1 restriction enzyme. The RFLP products were detected by 2% agorose gel electrophoresis tecnique. Our research examined that V600E mutation in BRAF gene on 51 patients. In result of this study 15 patients mutant (29.4%), 28 patients heterozygous (54.9%) and 8 patients wild (15.7%) were analysed. According to the preliminary results of our study, the BRAF V600E mutation may be a potential prognostic factor in the evaluation of thyroid nodules.
Chromosomal abnormalities play crucial role in first trimester spontaneous miscarriages. Microdeletion and duplication analysis may also important to determine etiology of abortions. It was aimed to detect the aneuploidies and microdeletion/duplications in aborted fetal materials in the current study. The study was performed between November 2014 and June 2015 in Research Hospital of Canakkale Onsekiz Mart University, C¸anakkale/Turkey. In a total of 50 spontaneously aborted fetal materials were genotyped in the current results. Genomic DNA were isolated from aborted fetal materials manually. DNA samples were used for QFPCR (Aneufast; 13, 18, 21, X and Y) and MLPA (MRC-Holland P095 and P245) genotyping. Results were analysed in capillary electrophoresis system. Results were analyzed with the Coffalyser and Gene Mapper programs. 13 (26%) triploidy and aneuploidies were detected by QF-PCR technique and that results are confirmed by MLPA (P095). In addition microdeletions at ZFY, DMD genes were detected in aborted materials with MLPA (P245) analysis. Microdeletions may be the primer cause of the abortion in the current cases. It is not possible to detect that microdeletions by QF-PCR technique. MLPA is a very fast and cheap method to detect deletions/duplications and various probe sets are available and useful for detecting chromosomal etiology of abortions. QF PCR has also advantages to detect Uniparental Disomy. Our previous study showed 3 UPD at 16 aborted fetal material. Using different tecniques in same laboratory will improve diagnostic capability.
http://dx.doi.org/10.1016/j.jbiotec.2015.06.263 A case of SRY positive 46, XX male with speaking disorder Ruslan Bayramov ∗ , Meltem Cerrah Günes¸, Yagut Erdem, Munis Dündar Department of Medical Genetics, Erciyes University, Kayseri, Turkey E-mail address:
[email protected] (R. Bayramov). Mental retardation and behavioural instability sometimes are caused by chromosomal disorders. 2 years old male was brought to
http://dx.doi.org/10.1016/j.jbiotec.2015.06.265
