9 Human Sperm Cells
129
1. Wash ejaculate, decondensate sperms Fig. 2. Outline of interphase
FISH from human sperm cells
2. Place sperms for RNase and Pepsin
pretreatment on slide
3. DenatureDNA probe and target DNA
on slide and hybridize them together.
3. Remove superfluous probe and
analyze signal constellation in cells
- RNase working solution (l00 ng/Ill): 1 III stock solution plus 99 III
2xSSC
Stock solution: dissolve 10 mg RNase A (Roche) in 1 ml2 x SSC, boil for
10 min and store in portions of 100 III in -20
D C
- Pepsin stock solution 10%: Dissolve 1 g pepsin (Fluka) in 10 ml aqua
bidest and store in portions of 50 III at -20
D C
- 2x SSC: 50 ml 20 x SSC plus 450 ml aqua bidest
129
1. Wash ejaculate, decondensate sperms Fig. 2. Outline of interphase
FISH from human sperm cells
2. Place sperms for RNase and Pepsin
pretreatment on slide
3. DenatureDNA probe and target DNA
on slide and hybridize them together.
3. Remove superfluous probe and
analyze signal constellation in cells
- RNase working solution (l00 ng/Ill): 1 III stock solution plus 99 III
2xSSC
Stock solution: dissolve 10 mg RNase A (Roche) in 1 ml2 x SSC, boil for
10 min and store in portions of 100 III in -20
D C
- Pepsin stock solution 10%: Dissolve 1 g pepsin (Fluka) in 10 ml aqua
bidest and store in portions of 50 III at -20
D C
- 2x SSC: 50 ml 20 x SSC plus 450 ml aqua bidest