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Fig. 3. Generating the Hand-null mutant by homologous
recombination. (A) Strategy for replacing the Hand-coding
region with the mini-white gene by ends-out homologous recombination.
(B) RT-PCR of the four mutant lines obtained from the screen showing no
detectable Hand transcript in homozygous mutant larvae. (C) In situ
hybridization using Hand probe (green) did not detect any
Hand transcripts in homozygous Hand mutants (lower panel),
which were distinguished from heterozygous Hand mutant embryos (upper
panel) by using anti-ß-Gal antibody (red) to detect the presence of the
balancer chromosome.