Microsatellite genotyping for genetic quality testing using sperm cells in the mouse.
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.
Katoh
Hideki
H
Institute for Experimental Animals, Hamamatsu University School of Medicine, Shizuoka, Japan.
Yoshino
Sayaka
S
Inui
Yumiko
Y
Honda
Sayaka
S
Takabayashi
Shuji
S
D000818:Animals; D015925:Cryopreservation; D004247:DNA / Q000235:genetics*; D005819:Genetic Markers; D016678:Genome; D005838:Genotype; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR / Q000235:genetics*; D018895:Microsatellite Repeats / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D011785:Quality Assurance, Health Care / Q000379:methods*; D013094:Spermatozoa / Q000145:classification / Q000166:cytology*; D013698:Templates, Genetic
D005819:Genetic Markers; D004247:DNA