Katoh Hideki, Yoshino Sayaka, Inui Yumiko, Honda Sayaka, Takabayashi Shuji
Institute for Experimental Animals, Hamamatsu University School of Medicine, Shizuoka, Japan.
Exp Anim. 2005 Jul;54(4):373-6. doi: 10.1538/expanim.54.373.
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 microl-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 microl), provides sufficient quantity of template DNA for genetic quality testing.
我们试图确定对一个微卫星标记进行基因分型所需的精子细胞数量。使用20微升规模的PCR，从约760个或更多精子细胞中提取的粗基因组DNA产生了足够量的PCR产物。我们还研究了非离子去污剂对粗精子基因组DNA提取的影响。用补充了非离子去污剂的裂解缓冲液提取的粗精子基因组DNA扩增得到的PCR产物显示出更清晰的条带。总之，我们的结果表明，一小部分冷冻精子，其体积小于原始体积的1/10（10微升），可为遗传质量检测提供足够量的模板DNA。
