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Screening males for mutations using frozen sperm and TILLING
Males
are screened through a complementary, reverse genetics method
employing a TILLING strategy
(t
argeting i nduced l ocal
l esions i n g enomes).
ENU F1 males are sacrificed, tissue from the heart taken, and
their sperm cryopreserved. In a collaboration with Dr. Derek
Stemple's lab at the Wellcome Trust Sanger Institute in England
(http://www.sanger.ac.uk/Teams/Team31/geneticscreens.shtml),
the samples are sequenced at several different genes of interest
to look for mutant variants of the coding sequence. Please see
Dr. Stemple's website for more information on this method or
refer to the publication from the Stemple and Zimmerman labs1.
(http://www.sanger.ac.uk/Teams/Team31/reversescreen.shtml
)
1Goda
T, Abu-Daya A, Carruthers C, Clark MD, Stemple DL, et al. (2006)
Genetic screens for mutations affecting development of Xenopus
tropicalis . PloS Genet 2(6):e91.
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