Animals
We will be using
Lytechinus variegatus from Florida. The gametes and
embryos can be kept at room temperature.
Figure 1. Lytechinus variegatus
adults
Protocol
This experiment will
involve the segregation of eggs into four samples. Sample 1
will be the control, exposed to NO radiation. In
sample 2, the eggs will be irradiated before fertilization.
Sample 3 will be irradiated 10 minutes after fertilization.
Sample 4 will be irradiated 30 minutes after fertilization.
The experiment is to be carried out as follows.
Collection of Gametes
Follow procedure given in basic
protocol.
UV Radiation
Irradiate sample #2 (unfertilized eggs) using the UV
crosslinker set on the default setting of intensity 120,000
microJoules/cm2
for a time of two minutes. The crosslinker has radiation
with a wavelength of 250nm.
Fertilization
Fertilize all unirradiated samples as one batch, and
then split into three separate test tubes following the
completion of fertilization. Fertilize the irradiated sample
as a separate batch. For fertilization procedure, follow
instructions given in basic
protocol.
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