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Genetics in neonatal surgical practice
Published in Prem Puri, Newborn Surgery, 2017
DNA generated by PCR can be used in many different ways to detect an abnormality in the sequence. If the test is aimed at detecting a known sequence abnormally, such as the common 3 base pair deletion on the cystic fibrosis gene Phe508del, the PCR product can be analyzed using mutation-specific oligonucleotide primers, or a DNA restriction enzyme test. If the search is for an unknown DNA mutation, such as those seen in hereditary breast and ovarian cancer, then many pieces of DNA generated by PCR from the patient can have their sequence analyzed using a semiautomated DNA analyzer.
Evaluation of JUN, FN1 and LAMB1 polymorphisms in pterygium in a Chinese Han population
Published in Ophthalmic Genetics, 2022
Xiying Wu, Shiqi Dong, Yuting Xu, Ge Zhu, Ming Yan
Venous blood samples of patients and controls were harvested. Genomic DNA was extracted from the peripheral blood leukocytes using standard phenol/chloroform extraction. DNA was quantified by the NanoDrop-2000C (Thermo Fisher Scientific, United States) and stored at −20°C until use. Genotypes were determined by SNaPshot® (Applied Biosystems, Foster City, CA, USA) minisequencing method. The PCR products were purified and sequenced on an Applied Biosystems 3730xl DNA Analyser. GeneMapper4.0 was used to analyze the sequencing results. The accuracy of the genotyping was verified by randomized blind repeated testing of 10% samples.
Relationship between MTHFR Gene Polymorphisms and Gastrointestinal Tumors Development: Perspective from Eastern Part of Turkey
Published in Journal of Investigative Surgery, 2022
Ersoy Öksüz, Gökhan Görgişen, Gökhan Oto, Hülya Özdemir, Abbas Aras, Murat Öksüz, İsmail Musab Gülaçar, Muhammet Hamdi Demirkol
PCR products were purified by the ExoSap reaction and the Big dye-terminator sequencing kit (Applied Biosystems, CA, USA) was used for the primers during amplification. Sequencing fragments were detected by capillary electrophoresis using the ABI Prism 3130 DNA analyzer (Applied Biosystems). Sequence chromatograms were analyzed by Finch TV (Figures 1,2).