کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
3934166 1253370 2009 5 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
An efficient and reliable DNA extraction method for preimplantation genetic diagnosis: a comparison of allele drop out and amplification rates using different single cell lysis methods
موضوعات مرتبط
علوم پزشکی و سلامت پزشکی و دندانپزشکی زنان، زایمان و بهداشت زنان
پیش نمایش صفحه اول مقاله
An efficient and reliable DNA extraction method for preimplantation genetic diagnosis: a comparison of allele drop out and amplification rates using different single cell lysis methods
چکیده انگلیسی

ObjectiveTo evaluate methods of DNA extraction from single cells for their suitability to amplify and provide a correct diagnosis of target disease genes.DesignExperimental study.SettingUniversity hospital laboratory.Patient(s)Two normal adult male and female blood donors.Intervention(s)Exon 51 of the dystrophin gene and the ZFX/ZFY gene were amplified from single lymphocytes using nested PCR. Five different methods of DNA extraction were tested (lysis in distilled water with freezing and thawing using liquid nitrogen, lysis in distilled water, alkaline lysis buffer, Proteinase K/sodium dodecyl sulfate (SDS) buffer, and N-lauroylsarcosine salt solution).Main Outcome Measure(s)Allele drop out and amplification rate.Result(s)The amplification efficiency from single unaffected lymphocytes was 89.0% using the liquid nitrogen method, 88.1% with the distilled water lysis method, 97.5% with the alkaline lysis buffer method, 91.5% with the Proteinase K/SDS lysis buffer method, and 84.8% using the N-lauroylsarcosine salt solution method. The mean allele drop out rate was 16.7%, 43.9%, 2.0%, 9.8%, and 18.9%, respectively, for each lysis method using single male lymphocytes as a template.Conclusion(s)Based on these results, DNA extraction using an alkaline lysis buffer results in more efficient rates of DNA amplification and less allele drop out than the other methods of DNA extraction tested. This method is suitable for the lysis of single cells in clinical preimplantation genetic diagnosis.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Fertility and Sterility - Volume 92, Issue 2, August 2009, Pages 814–818
نویسندگان
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