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Fertilization
- The eggs were washed several times with ASW by
allowing them to settle and pouring off the water. This
will removed some of the jelly coat.
- Enough eggs to cover the bottom of a 50 ml beaker
were transfered along with 10 ml of 10mM p-aminobenzoic
acid in ASW.
- 50 ml of sperm were diluted in 5 ml of ASW. 2 drops
were used to fertilize eggs.
- The presence of the fertilization envelope was
checked after 5 and 10 minutes in the control
culture.
- The eggs were poured through a nylon filter mounted
to the end of a 50 ml beaker. This was repeated until
most of the fertilization envelopes have been
removed.
- The cells were allowed to settle for a few
minutes.
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