Investigating the usage of molted feather samples as a DNA source with two methods in gender ıdentification of african grey parrot (Psittacus erithacus) by molecular analyses of CHDW and CHDZ genes

Bu çalışmanın amacı Afrika gri papağanlarında (Psittacus erithacus) cinsiyet tayininde kullanmak üzere dökülmüş tüylerden DNA izolasyonunda iki metodun (metot 1 ve metot 2) etkinliğinin değerlendirilmesidir. Oniki tanesi üç aya kadar saklanmış; iki tanesi dışkıyla kontamine, ayrıca sekiz tanesi beş aya kadar saklanmış; biri dışkıyla kontamine olmak üzere toplam yirmi papağana ait dökülmüş tüy örnekleri kullanılmıştır. Üç aya kadar saklanmış ve dışkıdan ari on adet dökülmüş tüyden metot 1 ile genomik DNA başarı ile elde edilmiştir. Cinsiyet ayırımı kromo helikaz-bağlanma bölgesi genlerinin (CHDW ve CHDZ) çoğaltılması ile iki dişi sekiz erkek olarak sonuçlanmıştır. Ancak genomik DNA metot 1 ile dışkı ile kontamine olmuş tüyler ile birlikte beş aya kadar saklanmış olan tüylerin hiçbirinden, ayrıca metot 2 ile tüy örneklerinin tamamından elde edilememiştir. Dökülmüş tüylerin dışkı ile kontaminasyonu ve tazeliği cinsiyet tayininin başarısını etkilemektedir. Sonuç olarak metot 1, Afrika gri papağanlarında temiz ve taze dökülmüş tüylerden DNA izolasyonu yapılarak cinsiyet tayininde kullanılabilir.The aim of this study was to evaluate the efficiency of two methods (method 1 and 2) for DNA isolation from molted feathers that were used in gender identification of African grey parrots (Psittacus erithacus). The molted feathers of twelve parrots were stored up to three months, two were contaminated with feces, further eight were stored up to five months; one was contaminated with feces totally feather samples of twenty parrots were used. Genomic DNA was isolated with method 1 successfully from the molted feathers of ten parrots that were stored for up to three months and free from feces. The differentiation of the gender that was made by amplification of chromo helicase-binding domain genes (CHDW and CHDZ), was resulted with two females and eight males. However no genomic DNAs were obtained from the feathers contaminated with feces or were stored up to five months with method 1 and none of the samples that were processed with method 2. Feces contamination and freshness of molted feathers affect the gender identification. In conclusion method 1 can be used in DNA isolation in order to perform gender identification from clean and fresh molted feathers in African grey parrots

Genetic Variability of CAST Gene in Native Sheep Breeds of Turkey

The aim of this study is to determine the genetic variability of CAST gene in native sheep breeds of Turkey by PCR-RFLP method. Six different native sheep breeds; Kivircik, Imroz, Karayaka, Hemsin, Red Karaman and Karakul were used in this study. This study was the first report about CAST gene variation in Karayaka, Red Karaman and Hemsin sheep breeds. After DNA isolation and PCR amplification, RFLP was performed with MspI enzyme. Two alleles M (336bp and 286bp) and N (622bp) were identified on 2% agarose gel electrophoresis. Allel and genotype frequencies, observed (Ho) and expected heterozygosity (He) and deviation from Hardy Weinberg Equilibrium were estimated by statistical analyses. The frequency of M allele was highest in Imroz (96%) and N allele was identified most frequently in Kivircik (30%) breed. Highest frequencies of MN genotype were identified in Kivircik (60%), MM in Imroz (92.6%) and NN in Red Karaman (7.1%) breeds respecitvely. Kivircik, Imroz, Karayaka and Karakul breeds were null from NN genotype. Kivircik sheep showed the highest heterozygosity (60%) and Imroz had the lowest (7.4%). The highest heterozygosity value was identified in Kivircik (60%), the lowest in Imroz (7.4%). All breeds except Kivircik and Hemsin were found in Hardy-Weinberg equilibrium. Absence of NN genotype in some breeds and high frequency of MN genotype in Kivircik breed might be resulted from the selection process of native sheep breeds in their breeding regions

Determining Genetic Variation of Calpastatin Gene with Mspl and Ncol Enzymes by Using PCR-RFLP Method in Kivircik Lambs

The aim of this study was to determine the genetic variation of calpastatin gene by using PCR-RFLP method with Mspl and Ncol enzymes in Kivircik lambs. Blood samples of Kivircik lambs (n=153) that were collected from eight different farms located in Kirklareli province, were used for DNA isolation. After PCR amplification, products were digested with Mspl and Ncol enzymes to differentiate M and N alleles on 2% and 3% agarose gel electrophoresis. The frequency of M and N alleles were found 88.2% and 11.8% for Mspl locus and 98.7% and 1.3% for Ncol locus respectively. The frequencies of MM, MN and NN genotypes were identified 0.77, 0.22 and 0.01 respectively for Mspl locus. The frequencies of MM and MN genotypes were determined 0.97 and 0.03 respectively for Ncol locus, NN genotype was not observed. Observed heterozygosity was higher in Mspl (0.22) than Ncol locus (0.03). Mspl and Ncol loci were found in Hardy-Weinberg equilibrium in Kivircik lambs raised in Kirklareli province