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|Title||Amplification of X- and Y-Chromosome-Specific Regions from Single Human Blastomeres by Polymerase Chain Reaction|
The major technological hurdle for preimplantation genetic diagnosis (PGD) is the inability to provide sufficient quantity and quality of DNA from single blastomeres. Therefore, much effort has been invested in developing methods to provide sufficient quantity of DNA for downstream molecular analyses. The whole genome amplification (WGA) is one approach designed to overcome these problems. This technique would amplify all of the genetic material present in a sample containing low amounts of DNA such as that present in a single cell. The product of WGA method should be amenable for use in polymerase chain reaction (PCR), Real-time PCR (RT-PCR) and comparative genomic hybridization (CGH) - based genetic analysis of single cell samples, such as blastomeres, and for testing of aneuploidy, single gene defects and sex selection. The objective of this study was to apply WGA and PCR techniques to sex determination of preimplantation human embryos (that were created by intracytoplasmic sperm injection "ICSI") for the first time in Gaza Strip. In this study, parts of SRY (Sex-Determining Region on Y chromosome) and glucose-6- phosphate dehydrogenase (G6PD) genes were amplified in 30 donated surplus embryos by PCR techniques. This study proved that WGA using the REPLI-g Mini Kit (Qiagen,USA) is applicable for very low amounts of DNA and this would open the door to diagnose several inherited diseases by genetic analysis to prevent couples who have risk for passing an inherited disease to offspring from giving birth to affected offspring. Sex determination by PCR is a fast and convenient technique, requiring only one PCR reaction, with the products being easily visualized on an agarose gel and it is useful for examining the sex of embryos prior to transfer to mother′s uterus.
|Publisher||الجامعة الإسلامية - غزة|
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