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Microsatellite Genotyping for Genetic Quality Testing Using Sperm Cells in the Mouse

机译:微卫星基因分型用于小鼠精子细胞遗传质量检测

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摘要

We attempted to determine the number of sperm cells required for genotyping of one microsatellite marker. The crude genomic DNA extracted from about 760 or more sperm cells gave sufficient quantity of PCR product using a 20 μl-scale PCR. We also studied the effects of non-ionic detergents on extraction of crude sperm genomic DNA. PCR products amplified with the crude sperm genomic DNA extracted using the lysis buffer supplemented with non-ionic detergents showed much clear bands. In conclusion, our results suggest that a small part of the frozen sperm, which is less than 1/10 of the original volume (10 μl), provides sufficient quantity of template DNA for genetic quality testing.
机译:我们试图确定一种微卫星标记基因分型所需的精子细胞数量。使用20μl规模的PCR,从约760个或更多精子细胞中提取的粗基因组DNA产生了足够数量的PCR产物。我们还研究了非离子去污剂对精子精子基因组DNA提取的影响。用添加有非离子型去污剂的裂解缓冲液提取的粗精子基因组DNA扩增的PCR产物显示出许多清晰的条带。总之,我们的结果表明,冷冻精子的一小部分少于原始体积的10/10(10μl),可为遗传质量检测提供足够数量的模板DNA。

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